Method of preventing cochlear disease, disorder or condition using (+)-azasetron

ABSTRACT

Disclosed is a method for treating ear disorders, including the administration of (+)-azasetron, or a pharmaceutically acceptable salt and/or solvate thereof.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a Continuation of application Ser. No. 16/093,911,filed on Oct. 15, 2018, which is the National Phase under 35 U.S.C. §371 of International Application No. PCT/EP2017/059058, filed on Apr.14, 2017, which claims the benefit under 35 U.S.C. § 119(e) to U.S.Provisional Application No. 62/322,690, filed on Apr. 14, 2016, andunder 35 U.S.C. § 119(a) to patent application Ser. No. 16/180,192.3,filed in Europe on Jul. 19, 2016, all of which are hereby expresslyincorporated by reference into the present application.

FIELD OF INVENTION

The present invention relates to the field of ear disorders, inparticular to methods for treating ear disorders. The present inventionrelates to (+)-azasetron, or pharmaceutically acceptable salts and/orsolvates thereof, for use in the treatment of an ear disorder.

BACKGROUND OF INVENTION

In 2013, the world health organization estimated about 360 millionpersons (over 5% of the world's population) affected by an ear disorderdue to diverse causes: noise, genetic conditions, complications atbirth, certain infectious diseases, chronic ear infections, the use ofparticular drugs, and ageing. This number will increase with ageingpopulation in the western countries and teenagers or young adults underincreased exposure to unsafe level of noise.

The ear is comprised of three major structural components: the outer,middle, and inner ears, which function together to convert sound wavesinto nerve impulses that travel to the brain, where they are perceivedas sound. The inner ear also helps to maintain balance.

The middle ear consists of the eardrum and a small air-filled chambercontaining a sequence of three tiny bones known as the ossicles, whichlink the eardrum to the inner ear. The inner ear (labyrinth) is acomplex structure consisting of the cochlea, which is the organ ofhearing and the vestibular system, the organ of balance. The vestibularsystem consists of the saccule, the utricle, and the semicircularcanals, which all help determine position and acceleration of the headin space and thus maintain balance.

The cochlea houses the organ of Corti, which consists, in part, of about20,000 specialized sensory cells, called “inner ear hair cells” or “haircells”. These cells have small hairline projections (cilia) that extendinto the cochlear fluid. Sound vibrations transmitted from the ossiclesin the middle ear to the oval window in the inner ear cause the fluidand cilia to vibrate. Hair cells in different parts of the cochleavibrate in response to different sound frequencies and convert thevibrations into nerve impulses which are sent to the brain forprocessing and interpretation. The inner ear hair cells are surroundedby inner ear support cells. Supporting cells underlie, at leastpartially surround, and physically support sensory hair cells within theinner ear. Representative examples of supporting cells include inner rod(pillar cells), outer rod (pillar cells), inner phalangeal cells, outerphalangeal cells (of Deiters), cells of Held, cells of Hensen, cells ofClaudius, cells of Boettcher, interdental cells and auditory teeth (ofHuschke).

The spiral ganglion is the group of nerve cells that send arepresentation of sound from the cochlea to the brain. The cell bodiesof the spiral ganglion neurons are found in the spiral structure of thecochlea and are part of the central nervous system. Their dendrites makesynaptic contact with the base of hair cells, and their axons arebundled together to form the auditory portion of the eighth cranialnerve (vestibulocochlear nerve).

Similarly, sensory hair cells in the vestibule respond to movements ininner ear fluids induced by head movements. The sensory hair cellresponses are converted into nerve impulses in vestibular ganglionneurons through dendritic synaptic contacts from neurons to the base ofvestibular hair cells. The vestibular neurons send representation ofhead position and movement in space to the brain through their axonbundles forming the vestibular portion of the vestibulocochlear nerve.

A variety of conditions may affect specifically one of these structuresof the ear impairing hearing or balance: ear infections are the mostcommon illness in infants and young children; tinnitus, a roaring in theears, can be the result of loud noises, medicines or a variety of othercauses; Meniere's disease may be the result of fluid problems in theinner ear, its symptoms include tinnitus and dizziness; ear barotraumais an injury to the ear because of changes in barometric (air) or waterpressure. Moreover, hearing loss and vertigo may be induced bydrug-based ototoxicity, infections, trauma, ischemia or congenitalcauses. Such conditions may affect different parts of the ear.

Various pharmaceutical compounds have already been tested for thetreatment of ear diseases: antibiotics, anti-inflammatory agents, orNMDA antagonists for example. Despite all the compounds tested, thesedrugs either exert strong adverse events, lack of efficacy, show anefficacy in the early symptoms only but no efficacy on chronic basis orshow an efficacy on a small cohort of patients only. Therefore, there isa need to discover novel treatment for ear diseases that could be usedfor a wild range of persons.

In particular, the application WO2010/133663 provides evidence ofcompounds from the setron family including azasetron, for the treatmentof lesional vestibular disorders.

The present application evidences that the enantiomer (+)-azasetron, ora pharmaceutically acceptable salt and/or solvate thereof, providessurprisingly good local drug exposure and results for the treatment ofear disorders, compared to racemic azasetron or enantiomer(−)-azasetron.

SUMMARY

One object of the invention is (+)-azasetron, of Formula (R)-I:

or a pharmaceutically acceptable salt and/or solvate thereof, for use inthe treatment of an ear disorder.

Another objection of the invention is a composition for use in thetreatment of an ear disorder comprising (+)-azasetron, of Formula (R)-I:

or a pharmaceutically acceptable salt and/or solvate thereof;

and at least one pharmaceutically acceptable excipient.

In one embodiment, the pharmaceutically acceptable salt is selected from(+)-azasetron besylate, (+)-azasetron malate and (+)-azasetronhydrochloride, and preferably is (+)-azasetron besylate.

In one embodiment, the composition is to be administered at a doseranging from about 0.01 mg to about 100 mg of (+)-azasetron. In anotherembodiment, the composition is to be administered at a dose ranging fromabout 0.1 mg to about 100 mg of (+)-azasetron.

In one embodiment, the composition is to be administered systemically orlocally, preferably systemically, more preferably orally.

In one embodiment, the ear disorder is selected from the groupcomprising: inner ear disorder, middle ear disorder, external eardisorder, infection of the ear, inflammation of the ear or autoimmunedisorder of the ear. In one embodiment, the ear disorder is selectedfrom hearing loss, tinnitus and a vestibular disorder.

In one embodiment, hearing loss is sensorineural hearing loss.

In one embodiment, sensorineural hearing loss is ototoxiccompound-induced hearing loss, preferably platinum-induced hearing loss,more preferably cisplatin-induced hearing loss.

In one embodiment, the composition is to be administered to a subjectdiagnosed with cancer. In one embodiment, the subject diagnosed withcancer is awaiting the receipt of, or is receiving platinum-basedchemotherapy, preferably cisplatin, carboplatin, oxaliplatin or acombination thereof. In one embodiment, the composition is to beadministered before, during and/or after platinum-based chemotherapy,preferably cisplatin-, carboplatin- or oxaliplatin-based chemotherapy ora combination thereof.

In one embodiment, sensorineural hearing loss is sudden sensorineuralhearing loss or noise-induced hearing loss.

In one embodiment, vestibular disorder is lesional vestibular disorder.

In one embodiment, the composition is an immediate release compositionor a sustained release composition. In one embodiment, the compositionis an orodispersible composition.

Definitions

In the present invention, the following terms have the followingmeanings:

-   -   “About” preceding a figure means plus or less 10% of the value        of said figure.    -   “Pharmaceutically acceptable excipient” refers to an excipient        that does not produce any adverse, allergic or other unwanted        reactions when administered to an animal, preferably a human. It        includes any and all solvents, dispersion media, coatings,        antibacterial and antifungal agents, isotonic and absorption        delaying agents and the like. For human administration,        preparations should meet sterility, pyrogenicity, general        safety, quality and purity standards as required by regulatory        offices, such as, for example, FDA Office or EMA.    -   “Subject” refers to a mammal, preferably a human. In one        embodiment, the subject is a pet, including, without limitation,        a dog, a cat, a guinea pig, a hamster, a rat, a mouse, a ferret,        a rabbit, a bird or an amphibian. In one embodiment, a subject        may be a “patient”, i.e., a female or a male, an adult or a        child, who/which is awaiting the receipt of, or is receiving        medical care or was/is/will be the object of a medical        procedure, or is monitored for the development of an ear        disease, disorder or condition.    -   “Therapeutically effective amount” refers to level or amount of        agent that is aimed at, without causing significant negative or        adverse side effects to the target, (1) delaying or preventing        the onset of an ear disease, disorder, or condition; (2) slowing        down or stopping the progression, aggravation, or deterioration        of one or more symptoms of the ear disease, disorder, or        condition; (3) bringing about ameliorations of the symptoms of        the ear disease, disorder, or condition; (4) reducing the        severity or incidence of the ear disease, disorder, or        condition; or (5) curing the ear disease, disorder, or        condition. A therapeutically effective amount may be        administered prior to the onset of the ear disease, disorder, or        condition, for a prophylactic or preventive action.        Alternatively or additionally, the therapeutically effective        amount may be administered after onset of the ear disease,        disorder, or condition, for a therapeutic action. In one        embodiment, a therapeutically effective amount of the        composition is an amount that is effective in reducing at least        one symptom of an ear disease, disorder or condition.    -   “Treating” or “treatment” or “alleviation” refers to both        therapeutic treatment and prophylactic or preventative measures;        wherein the object is to prevent or slow down (lessen) the        targeted pathologic condition or disorder. Those in need of        treatment include those already with the disorder as well as        those prone to have the disorder or those in whom the disorder        is to be prevented. A subject or mammal is successfully        “treated” for the targeted pathologic disorder if, after        receiving a therapeutic amount of the compound or composition of        the present invention, the patient shows observable effects on        one or more of the followings; (i) promotion of ear        functioning; (ii) decrease in hearing loss; (iii) relief to some        extent, of one or more of the symptoms associated with the        specific disorder or condition, such as vertigo, dizziness and        tinnitus; (iv) reduced morbidity and mortality, and (v)        improvement in quality of life issues. The above parameters for        assessing successful treatment and improvement in the disorder        are readily measurable by routine procedures familiar to a        physician.    -   “Prodrug” refers to the pharmacologically acceptable derivatives        of compounds of Formula I of the invention, such as for example        esters, whose in vivo biotransformation product generates the        biologically active drug. Prodrugs are generally characterized        by increased bio-availability and are readily metabolized into        biologically active compounds in vivo.    -   “Predrug” refers to any compound that will be modified to form a        drug species, wherein the modification may take place either        inside or outside of the body, and either before or after the        predrug reaches the area of the body where administration of the        drug is indicated.    -   “Solvate” is used herein to describe a molecular complex        comprising the compound of Formula I of the invention and one or        more pharmaceutically acceptable solvent molecules, for example,        ethanol. The term “hydrate” is employed when said solvent is        water.    -   “Bioavailability” refers to the rate and extent to which a drug        or other substance becomes available to the target tissue (i.e.,        reaches the target tissue), e.g., the inner ear, after        administration.    -   “Vestibular disorders” refers to a vast family of conditions        wherein the vestibular organ is associated. These disorders may        be distinguished by their putative origins, one can thus        identify (1) lesional vestibular disorders and (2) non lesional        vestibular disorders.    -   “Lesional vestibular disorders” refer to vestibular disorders        wherein lesions of inner ear cells and/or vestibular nerve are        present or will appear during the disorder time course. In this        case, the functionality of the vestibule is impaired. However,        morphofunctional alterations of the vestibular endorgans cannot        be evaluated directly (except for large lesions that can be        detected by MRI). Conversely, indirect assessment methods are        currently used to evaluate the loss of functionality of the        vestibule. These testing methods are generally conducted at ENT        clinic/hospitals. Among them we can cite the videonystagmography        (VNG) and assessment of the vestibulo-ocular reflex (VOR) using        caloric or rotational tests, video head impulse testing (vHIT)        and vestibular evoked myogenic potentials (VEMP). Lesional        vestibular disorders include:        -   vestibular disorders wherein an inflammation of the inner            ear and/or the vestibular nerve induces reversible and/or            irreversible damages. Examples of conditions from this group            include, but are not limited to, vestibular neuritis, acute            unilateral vestibulopathy and vestibular neuronitis;        -   vestibular disorders wherein inner ear fluids are affected            (abnormalities in the quantity, composition, and/or pressure            of the endolymph), these disorders usually develop lesions            during the disease time course. Examples of conditions from            this group are Meniere's disease and secondary endolymphatic            hydrops. They are associated with tinnitus and hearing loss;        -   vestibular disorders induced by insults or lesions of the            vestibular endorgans. Examples of said conditions are            vertigo caused by local ischemia, excitotoxicity, trauma            that affect temporal bones or ototoxic insult to vestibular            hair cells by drugs such as gentamicin and cisplatin;    -   iterative vestibular disorders of unknown origin leading to        permanent vestibular deficits, but without tinnitus or hearing        loss. An example of a condition from this group is vestibular        migraine (or migrainous vertigo).    -   “Non-lesional vestibular disorders” refer to vestibular        disorders supported by transient and often iterative vertigo        crisis wherein no lesion on inner ear cells and/or vestibular        nerve can be observed. In this case, the functionality of the        vestibule evaluated between the vertigo crisis using functional        tests (VOR, VNG) does not differ from healthy vestibule.        Non-lesional vestibular disorders include:        -   vestibular disorders wherein debris had been collected            within a part of the inner ear. This debris, called            otoconia, is made up of small crystals of calcium carbonate            and when they shift, they send false signals to the brain.            Examples of said conditions include, but are not limited to,            positional vertigos and in particular benign paroxysmal            positional vertigo (BPPV);        -   iterative vestibular disorders of unknown origin without            tinnitus, hearing loss or permanent vestibular deficits.

DETAILED DESCRIPTION

One object of the invention is a compound of Formula I:

or a pharmaceutically acceptable salt and/or solvate thereof, wherein *stands for the (R)-enantiomer (corresponding to the (+)-enantiomer), the(S)-enantiomer (corresponding to the (−)-enantiomer), the racemate or anon-racemic mixture of (R) and (S) enantiomers (corresponding tomixtures of (+)- and (−)-enantiomers).

Compound of Formula I is called “azasetron” and corresponds to6-chloro-3,4-dihydro-N-(8-methyl-8-azabicyclo-[3.2.1]-oct-3-yl)-2,4-dimethyl-3-oxo-2H-1,4-benzoxazine-8-carboxamide,which may also be referred to asN-(1-azabicyclo[2.2.2]octan-8-yl)-6-chloro-4-methyl-3-oxo-1,4-benzoxazine-8-carboxamide,as described in the U.S. Pat. No. 4,892,872, which is incorporatedherein by reference.

Azasetron has one chiral center that can give rise to two stereoisomers.

In one embodiment, the invention refers to racemic azasetron, or apharmaceutically acceptable salt and/or solvate thereof.

In one embodiment, the invention refers to the (R)-enantiomer ofazasetron, or a pharmaceutically acceptable salt and/or solvate thereof.The (R)-enantiomer of azasetron is called (R)-azasetron or(+)-azasetron, corresponding to Formula (R)-I:

In one embodiment, the invention refers to the (S)-enantiomer ofazasetron, or a pharmaceutically acceptable salt and/or solvate thereof.The (S)-enantiomer of azasetron is called (S)-azasetron or(−)-azasetron, corresponding to Formula (S)-I:

In a particularly preferred embodiment, the invention relates to(+)-azasetron, or a pharmaceutically acceptable salt and/or solvatethereof. Indeed, the Applicant surprisingly demonstrates that theadministration of (+)-azasetron results in higher local exposure in theinner ear than racemic azasetron or (−)-azasetron. Therefore,bioavailability in the ear of (+)-azasetron is greater than that ofracemic azasetron. In one embodiment, bioavailability in the ear of(+)-azasetron is at least 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9,2.0 or more times greater than that of racemic azasetron and/or(−)-azasetron.

The compounds of the invention include compounds of Formula I,preferably of Formula (R)-I as hereinbefore defined, including allpolymorphs and crystal habits thereof, predrugs and prodrugs thereof andisotopically-labeled compounds of Formula I, preferably of Formula(R)-I.

The compounds of the invention may be in the form of pharmaceuticallyacceptable salts. Pharmaceutically acceptable salts of the compounds ofFormula I include the acid addition salts thereof. Suitable acidaddition salts are formed from acids which form non-toxic salts.Examples of acid addition salts include the besylate,hydrochloride/chloride, malate, benzoate, ethane-1,2-disulfonate,fumarate, tartrate, acetate, adipate, ascorbate, aspartate,bicarbonate/carbonate, bisulphate/sulphate, borate, camsylate, citrate,cyclamate, edisylate, esylate, ethanesulfonate, formate, gluceptate,gluconate, glucuronate, glutamate, hexafluorophosphate, hibenzate,hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, maleate,malonate, mesylate, methylsulphate, naphthylate, 2-napsylate,nicotinate, nitrate, orotate, oxalate, palmitate, pamoate,phosphate/hydrogen phosphate/dihydrogen phosphate, pyroglutamate,saccharate, stearate, succinate, tannate, p-toluenesulfonate, tosylate,trifluoroacetate and xinofoate salts.

In one embodiment, preferred acid addition salts include besylate,hydrochloride, malate, benzoate, ethane-1,2-disulfonate, fumarate andtartrate salts; more preferably besylate, hydrochloride, malate andbenzoate salts; even more preferably besylate, hydrochloride or malatesalts.

According to a specific embodiment, the invention relates to(+)-azasetron besylate, (+)-azasetron hydrochloride, (+)-azasetronmalate, (+)-azasetron benzoate, (+)-azasetron ethane-1,2-disulfonate,(+)-azasetron fumarate and (+)-azasetron tartrate. According to apreferred embodiment, the invention relates to (+)-azasetron besylate,(+)-azasetron hydrochloride, (+)-azasetron malate, (+)-azasetronbenzoate; more preferably the invention relates to (+)-azasetronbesylate, (+)-azasetron hydrochloride or (+)-azasetron malate.

Pharmaceutically acceptable salts of compounds of Formula I may beprepared by reacting the compound of Formula I with the desired acid, orby converting one salt of the compound of Formula I to another byreaction with an appropriate acid or by means of a suitable ion exchangecolumn. All these reactions are typically carried out in solution.

The salt may precipitate from solution and be collected by filtration ormay be recovered by evaporation of the solvent. The degree of ionizationin the salt may vary from completely ionized to not ionized resulting ina co-crystal.

Examples of processes that may be used to synthetize (+)-azasetron or apharmaceutically acceptable salt and/or solvate thereof are well knownin the art. Significant literature and background information isavailable on the synthesis of azasetron. Reference can be made, e.g., toChinese patents CN101786963 and CN104557906.

In one embodiment, synthesis of (+)-azasetron or of a pharmaceuticallyacceptable salt and/or solvate thereof includes ab initio synthesisand/or chiral resolution.

In one embodiment, when ab initio synthesis of (+)-azasetron or of apharmaceutically acceptable salt and/or solvate thereof is implemented,at least one racemic starting compound and/or intermediate compound issubstituted by a chiral compound.

Another object of the invention is a composition comprising, consisting,or consisting essentially of at least one compound of Formula I or apharmaceutically acceptable salt and/or solvate thereof.

Another object of the invention is a pharmaceutical compositioncomprising, consisting, or consisting essentially of at least onecompound of Formula I or a pharmaceutically acceptable salt and/orsolvate thereof and at least one pharmaceutically acceptable excipient.

Another object of the invention is a medicament comprising, consisting,or consisting essentially of at least one compound of Formula I or apharmaceutically acceptable salt and/or solvate thereof.

In one embodiment, the composition, pharmaceutical composition ormedicament of the invention comprises, consists or consists essentiallyof (+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof, preferably (+)-azasetron besylate, (+)-azasetron hydrochlorideor (+)-azasetron malate.

As used herein, the term “consist essentially of”, with reference to acomposition, pharmaceutical composition or medicament, means that the atleast one compound of the invention is the only one therapeutic agent oragent with a biologic activity within said composition, pharmaceuticalcomposition or medicament.

In one embodiment, the composition, pharmaceutical composition ormedicament of the invention does not comprise of (−)-azasetron or apharmaceutically acceptable salt and/or solvate thereof. In oneembodiment, the composition, pharmaceutical composition or medicament ofthe invention comprises less than about 40% w/w, 30% w/w, 20% w/w or 10%w/w of (−)-azasetron or a pharmaceutically acceptable salt and/orsolvate thereof in weight to the total weight of azasetron, preferablyless than about 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% w/w (−)-azasetron ora pharmaceutically acceptable salt and/or solvate thereof in weight tothe total weight of azasetron, more preferably less than about 0.5%,0.4%, 0.3%, 0.25%, 0.2%, 0.15%, 0.1% w/w (−)-azasetron or apharmaceutically acceptable salt and/or solvate thereof in weight to thetotal weight of azasetron.

In one embodiment, the composition, pharmaceutical composition ormedicament of the invention does not comprise a mixture of (+)-azasetronand (−)-azasetron, or pharmaceutically acceptable salts and/or solvatesthereof.

In one embodiment, the composition, pharmaceutical composition ormedicament of the invention comprises at least a 60:40 w/w mixture,preferably at least a 70:30, 80:20 or 90:10 w/w mixture of(+)-azasetron:(−)-azasetron, or pharmaceutically acceptable salts and/orsolvates thereof, preferably at least a 95:5, 96:4, 97:3, 98:2, 99:1 w/wmixture of (+)-azasetron:(−)-azasetron, or pharmaceutically acceptablesalts and/or solvates thereof, more preferably a 99.5:0.5, 99.6:0.4,99.7:0.3, 99.75:0.25, 99.8:0.2, 99.85:0.15, 99.9:0.1 w/w mixture of(+)-azasetron:(−)-azasetron, or pharmaceutically acceptable salts and/orsolvates thereof.

Examples of pharmaceutically acceptable excipients include, but are notlimited to, water, saline, Ringer's solution, dextrose solution, andsolutions of ethanol, starch, glucose, sucrose, dextran, mannose,mannitol, sorbitol, polyethylene glycol (PEG), phosphate, acetate,gelatin, collagen, Carbopol©, vegetable oils, and the like. One mayadditionally include suitable preservatives, stabilizers, antioxidants,antimicrobials, and buffering agents, such as, for example, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), citric acid,ascorbic acid, tetracycline, and the like.

Carbopol© or “carbomer” refer to high molecular weight polymers ofpolyacrylic acid crosslinked with allyl sucrose or pentaerythritol allylether, said polymer comprising homopolymers and copolymers. Examples ofCarbopol® include, but are not limited to, Carbopol® 910, Carbopol® 934,Carbopol® 940, Carbopol® 941, Carbopol® 974, Carbopol® 981, Carbopol®Ultrez and Polycarbophil.

Other examples of pharmaceutically acceptable excipients that may beused in the composition of the invention include, but are not limitedto, ion exchangers, alumina, magnesium stearate, aluminum stearate,lecithin, serum proteins, such as human serum albumin, buffer substancessuch as phosphates, glycine, sorbic acid, potassium sorbate, partialglyceride mixtures of saturated vegetable fatty acids, water, salts orelectrolytes, such as protamine sulfate, disodium hydrogen phosphate,potassium hydrogen phosphate, sodium chloride, zinc salts, colloidalsilica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-basedsubstances, polyethylene glycol, sodium carboxymethylcellulose,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers,polyethylene glycol and wool fat.

In addition, pharmaceutically acceptable excipients may also comprise,without limitation, surfactants (e.g. hydroxypropylcellulose); suitablecarriers, such as, for example, solvents and dispersion mediacontaining, for example, water, ethanol, polyol (e.g. glycerol,propylene glycol, and liquid polyethylene glycol, and the like),suitable mixtures thereof, and vegetable oils, such as, for example,peanut oil and sesame oil; isotonic agents, such as, for example, sugarsor sodium chloride; coating agents, such as, for example,hydroxypropylmethylcellulose (HPMC), polyethylene glycol (PEG),polysorbate 80, titanium dioxide and lecithin; agents delayingabsorption, such as, for example, aluminum monostearate and gelatin;preservatives, such as, for example, benzalkonium chloride, benzethoniumchloride, chlorobutanol, thimerosal and the like; buffers, such as, forexample, boric acid, sodium and potassium bicarbonate, sodium andpotassium borates, sodium and potassium carbonate, sodium acetate,sodium biphosphate and the like; tonicity agents, such as, for example,dextrose, potassium chloride, propylene glycol, sodium chloride;antioxidants and stabilizers, such as, for example, sodium bisulfite,sodium metabisulfite, sodium thiosulfite, thiourea and the like;nonionic wetting or clarifying agents, such as, for example, polysorbate80, polysorbate 20, poloxamer 282 and tyloxapol; viscosity modifyingagents, such as, for example dextran 40, dextran 70, gelatin, glycerin,hydroxyethylcellulose, hydroxmethylpropylcellulose, lanolin,methylcellulose, petrolatum, polyethylene glycol, polyvinyl alcohol,polyvinylpyrrolidone, carboxymethylcellulose; diluents, adjuvants andthe like.

Another object of the invention is a compound, a composition, apharmaceutical composition or a medicament, as described hereinabove,for treating or for use in treating an ear disease, disorder orcondition.

The present invention thus relates to (+)-azasetron or apharmaceutically acceptable salt and/or solvate thereof (in particular(+)-azasetron besylate, (+)-azasetron hydrochloride, (+)-azasetronmalate or a mixture thereof) as described hereinabove, for treating, orfor use in treating an ear disease, disorder or condition.

In one embodiment, (+)-azasetron or a pharmaceutically acceptable saltand/or solvate thereof (in particular (+)-azasetron besylate,(+)-azasetron hydrochloride, (+)-azasetron malate or a mixture thereof)is for preventing an ear disease, disorder or condition. In anotherembodiment, (+)-azasetron or a pharmaceutically acceptable salt and/orsolvate thereof (in particular (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof) is foralleviating a symptom or for curing an ear disease, disorder orcondition.

The present invention also relates to a composition for treating an eardisease, disorder or condition, comprising or consisting of orconsisting essentially of (+)-azasetron or a pharmaceutically acceptablesalt and/or solvate thereof (in particular (+)-azasetron besylate,(+)-azasetron hydrochloride, (+)-azasetron malate or a mixture thereof).

In one embodiment, the composition of the invention is for preventing anear disease, disorder or condition. In another embodiment, thecomposition of the invention is for alleviating a symptom of an eardisease, disorder or condition or for curing an ear disease, disorder orcondition.

The present invention also relates to a pharmaceutical composition fortreating an ear disease, disorder or condition, comprising or consistingof or consisting essentially of (+)-azasetron or a pharmaceuticallyacceptable salt and/or solvate thereof (in particular (+)-azasetronbesylate, (+)-azasetron hydrochloride, (+)-azasetron malate or a mixturethereof) and at least one pharmaceutically acceptable excipient.

In one embodiment, the pharmaceutical composition of the invention isfor preventing an ear disease, disorder or condition. In anotherembodiment, the pharmaceutical composition of the invention is foralleviating a symptom of an ear disease, disorder or condition or forcuring an ear disease, disorder or condition.

The present invention also relates to a medicament for treating an eardisease, disorder or condition, comprising or consisting of orconsisting essentially of (+)-azasetron or a pharmaceutically acceptablesalt and/or solvate thereof (in particular (+)-azasetron besylate,(+)-azasetron hydrochloride, (+)-azasetron malate or a mixture thereof).

In one embodiment, the medicament of the invention is for preventing anear disease, disorder or condition. In another embodiment, themedicament of the invention is for alleviating a symptom of an eardisease, disorder or condition or for curing an ear disease, disorder orcondition.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating ear diseases, disordersor conditions selected from the group comprising, but not limited to,cochlear diseases, disorders or conditions; vestibular diseases,disorders or conditions; inner ear diseases, disorders or conditions;middle ear diseases, disorders or conditions; external ear diseases,disorders or conditions; infection of the ear; and inflammation of theear.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating cochlear diseases,disorders or conditions, including without limitation, hearing loss,tinnitus, cochlear ototoxicity (i.e., toxicity inducing damages of thecochlea), cochlear excitotoxic-inducing occurrence, head trauma withcochlear lesion, cochlear hyperacusis, and cochlear tinnitus.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating hearing loss, includingwithout limitation, sensorineural hearing loss, conductive hearing lossand mixed hearing loss (wherein mixed hearing loss is a combination ofboth sensorineural and conductive hearing loss).

Sensorineural hearing loss and conductive hearing loss may bedistinguished according to tests well known of the skilled artisan,including, without limitation, Weber and Rinne tests, otoscopy,audiograms, audiometry, auditory brainstem recordings, optoacousticemissions recordings, word recognition tests and speech-in-noise tests.

Methods for measuring hearing loss are well-known by the skilledartisan. Examples of such methods include, but are not limited to,tympanometry, acoustic reflex tests, tuning fork test, bone conductiontest, pure tone audiogram, behavioral observation audiometry, visualreinforcement audiometry, conditioned play audiometry, ABR (auditorybrainstem responses) measurement, DPOAE (distortion product otoacousticemissions) measurement, TEOAE (transiently evoked otoacoustic emissions)measurement, speech in noise test, word comprehension test and the like.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating sensorineural hearingloss including, without limitation, sudden sensorineural hearing loss(e.g., idiopathic sudden hearing loss), noise-induced sensorineuralhearing loss, drug-induced sensorineural hearing loss (e.g., ototoxiccompound-induced hearing loss), acquired sensorineural hearing loss,age-related hearing loss, congenital, hereditary and geneticsensorineural hearing loss (such as, but not limited to, Usher syndrome,Pendred syndrome, Jervell and Lange-Nielsen syndrome, Alport syndrome,Mohr-Tranebjaerg syndrome and Cogan syndrome) (i.e., sensory hair cells,neurons and supporting cells of the cochlea or the auditory nerve wereabnormal at birth or become abnormal during development) andsensorineural hearing loss induced by viral or bacterial infection (suchas, but not limited to, CMV).

Examples of causes of sensorineural hearing loss include, but are notlimited to, ototoxic compounds, excessive noise exposure (such as, forexample, exposure to a noise of more than about 70 dB, 80 dB, 90 dB, 100dB, 110 dB, 120 dB, 130 dB or more), aging, inflammation, inner earinvolvement by infectious agents (such as, for example, viral andbacterial infections), autoimmunity (such as, for example, autoimmuneinner ear disease) or vasculopathy, illnesses (including but not limitedto high blood pressure and diabetes), head trauma, tumors or blastexposure.

Examples of ototoxic compounds that may cause sensorineural hearing lossinclude, but are not limited to, chemotherapeutic agents (such as,without limitation, platinum drugs, including, without limitation,cisplatin, carboplatin, oxaliplatin, satraplatin, picoplatin,tetraplatin, transplatin, nedaplatin, ormaplatin, PtCl₂[R,RDACH],pyriplatin, ZD0473, BBR3464 and Pt-1C3), aminoglycosides (such as, butnot limited to, neomycin, gentamycin, kanamycin, amikacin andtobramycin), loop diuretics (such as, without limitation, furosemide,torsemide and bumetanide), macrolides (such as, without limitation,erythromycin and azithromycin), glycopeptides (such as, withoutlimitation, vancomycin) antimetabolites (such as, without limitation,methotrexate), antimalarials (such as, without limitation, quinine),antiretroviral drugs (such as, without limitation, abacavir, AZT,delavirdine, didenosine, efavirenz, emtricitabine, indinavir,lamivudine, nefinavir, nevirapine, tenofovir, ritonavir, stavudine andzalcitabine) and salicylates (such as, without limitation, aspirin).

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating ototoxic compound-inducedsensorineural hearing loss. In one embodiment, the compound,composition, pharmaceutical composition or medicament of the inventionis for treating chemotherapeutic agents-induced sensorineural hearingloss. In one embodiment, the compound, composition, pharmaceuticalcomposition or medicament of the invention is for treatingplatinum-induced sensorineural hearing loss, such as, withoutlimitation, sensorineural hearing loss induced by cisplatin,carboplatin, oxaliplatin, satraplatin, picoplatin, tetraplatin,transplatin, nedaplatin, ormaplatin, PtCl₂[R,RDACH], pyriplatin, ZD0473,BBR3464 and/or Pt-1C3, preferably cisplatin.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating compound-inducedototoxicity, including without limitation, ototoxicity induced bychemotherapeutic agents (such as, without limitation, platinum drugs,including, without limitation, cisplatin, carboplatin, oxaliplatin,satraplatin, picoplatin, tetraplatin, transplatin, nedaplatin,ormaplatin, PtCl₂[R,RDACH], pyriplatin, ZD0473, BBR3464 and Pt-1C3),aminoglycosides (such as, but not limited to, neomycin, gentamycin,kanamycin, amikacin and tobramycin), loop diuretics (such as, withoutlimitation, furosemide, torsemide and bumetanide), macrolides (such as,without limitation, erythromycin and azithromycin), glycopeptides (suchas, without limitation, vancomycin) antimetabolites (such as, withoutlimitation, methotrexate), antimalarials (such as, without limitation,quinine and analogs thereof), salicylates and analogs thereof (such as,without limitation, aspirin), radiation treatment, antibiotic agents,excitatory neurotoxins, immunosuppressive agents, β-blockers,vasodilator agents, anti-retroviral drugs (such as, without limitation,abacavir, AZT, delavirdine, didenosine, efavirenz, emtricitabine,indinavir, lamivudine, nefinavir, nevirapine, tenofovir, ritonavir,stavudine and zalcitabine), cardiovascular agents (e.g. ACE inhibitors),antiplatelet agents, radiocontrast media, immunoglobulins, mannitol,NSAIDs, heavy metals, lithium salts, catechol-O-methyltransferaseinhibitors, antiepileptic drugs, proton pump inhibitors,antidepressants, antihistamines (e.g. diphenhydramine, doxylamine),antibacterial sulfonamides, benzodiazepines, pesticides, neurotoxicagents (e.g. glutamate, Bicuculline, Botulinum toxin), alcohol, heavymetals, and drugs of abuse (e.g. cocain, heroin, ketamine). In oneembodiment, the compound, composition, pharmaceutical composition ormedicament of the invention is for treating radiation treatment-inducedototoxicity.

Examples of inflammatory causes of sensorineural hearing loss include,but are not limited to, autoinflammatory diseases (such as, for example,Muckle-Wells Syndrome), suppurative labyrinthitis, meningitis, mumps ormeasles.

Examples of viral causes of sensorineural hearing loss include, but arenot limited to, syphilis, mumps or measles.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating sudden hearing loss,preferably sudden sensorineural hearing loss (SSNHL).

Sudden hearing loss may be defined by the following audiometriccriterion: decrease in hearing of at least about 10 dB, 20 dB, 30 dB ormore over at least three contiguous frequencies, evolving within 3 daysor less. Because premorbid audiometry is generally unavailable, forunilateral hearing loss, hearing loss may be defined as relative to theopposite ear's thresholds or relative to population means.

Sudden hearing loss, preferably sudden sensorineural hearing loss(SSNHL), may result (without limitation) from vestibular schwannoma(acoustic neuroma), stroke, malignancy, vascular ischemia of the innerear, perilymph fistula or autoimmune causes (including, withoutlimitation, IgE or IgG allergy) or other causes (such as, for example,causes listed above). However, a cause of SSNHL is identified in only 10to 15% of patients at the time of presentation.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating acquired sensorineuralhearing loss, i.e., sensory hair cells, synapses and neurons of thecochlea or the auditory nerve were normal at birth, but weresubsequently damaged.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating congenital hearing loss,preferably congenital sensorineural hearing loss including, including,without limitation, a lack of development (aplasia) of the cochlea,chromosomal syndromes, hereditary hearing loss (such as, but not limitedto, Usher syndrome, Pendred syndrome, Jervell and Lange-Nielsensyndrome, Alport syndrome, Mohr-Tranebjaerg syndrome and Cogansyndrome), congenital cholesteatoma, delayed familial progressive,congenital rubella syndrome, and human Cytomegalovirus (HCMV)transmission to a developing fetus during pregnancy.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating conductive hearing loss,resulting from abnormalities of the external ear, tympanic membrane,middle ear space or ossicles. It may result from cerumen impaction,middle ear fluid, otitis media, foreign bodies, perforated tympanicmembrane, canal edema from otitis externa, otosclerosis, trauma (such asloss of residual hearing from electrode insertion trauma in cochlearimplant surgery or surgery-induced hearing loss) or cholesteatoma (allthese conditions may be diagnosed by otoscopy).

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating tinnitus. In oneembodiment, tinnitus is a symptom of hearing loss.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating vestibular diseases,disorders or conditions, including without limitation, lesionalvestibular disorders and non-lesional vestibular disorders.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating vestibular diseases,disorders or conditions, including without limitation, vestibularmigraine, vestibular neuritis (including, without limitation, vestibularneuronitis, viral neuronitis, labyrinthitis, viral endolymphaticlabyrinthitis, serous labyrinthitis, suppurative labyrinthitis), acuteunilateral vestibulopathy, vertigo and dizziness (including, withoutlimitation, migraine-associated vertigo, spontaneous episodic vertigo,benign positional paroxysmal vertigo, familial episodic vertigo,age-related dizziness and imbalance, motion sickness, mal dedebarquement), vestibular ototoxicity (including, without limitation,compound-induced and drug-induced ototoxicity, i.e., inducing impairmentof the vestibule function leading to vestibular deficits, and induced bycompounds listed without limitation above), vestibule-toxic impairments,hydrops (including, without limitation, endolymphatic hydrops, secondaryendolymphatic hydrops), Meniere's disease (including, withoutlimitation, spell of Meniere's disease, chronic Meniere disease),fistula (including, without limitation, perilymphatic fistula,labyrinthine fistula), trauma (including, without limitation, headtrauma with labyrinthine haemorrhage, barotrauma), infections(including, without limitation, chronic or acute labyrinthineinfection), autoimmune inner ear disease, benign or malignant tumors(including, without limitation, vestibular schwannomas, acousticneuroma), presbyvestibula, vestibular syndromes after chirurgicaltreatments of middle ear, channelopathies, superior semicircular canaldehiscence, endolymphatic sac or pontocerebellar angle, ataxia(including, without limitation, episodic ataxia), enlarged vestibularaqueduct, bilateral vestibular hypofunction, neurotoxic vestibulopathy,pediatric vestibular disorder, Cogan syndrome, vestibular hyperacusis,vertebrobasilar insufficiency.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating lesional vestibulardiseases, disorders or conditions, including without limitation,vestibular migraine, vestibular neuritis (including, without limitation,vestibular neuronitis, viral neuronitis, labyrinthitis, viralendolymphatic labyrinthitis, serous labyrinthitis, suppurativelabyrinthitis), acute unilateral vestibulopathy, vertigo and dizziness(including, without limitation, migraine-associated vertigo, spontaneousepisodic vertigo, familial episodic vertigo, age-related dizziness andimbalance), vestibular ototoxicity (including, without limitation,compound-induced and drug-induced ototoxicity, i.e., inducing impairmentof the vestibule function leading to vestibular deficits, and induced bycompounds listed without limitation above), vestibule-toxic impairments,hydrops (including, without limitation, endolymphatic hydrops, secondaryendolymphatic hydrops), Meniere's disease (including, withoutlimitation, spell of Meniere's disease, chronic Meniere disease),fistula (including, without limitation, perilymphatic fistula,labyrinthine fistula), trauma (including, without limitation, headtrauma with labyrinthine haemorrhage, barotrauma), infections(including, without limitation, chronic or acute labyrinthineinfection), autoimmune inner ear disease, benign or malignant tumors(including, without limitation, vestibular schwannomas, acousticneuroma), presbyvestibula, vestibular syndromes after chirurgicaltreatments of middle ear, channelopathies, superior semicircular canaldehiscence, endolymphatic sac or pontocerebellar angle, ataxia(including, without limitation, episodic ataxia), enlarged vestibularaqueduct, bilateral vestibular hypofunction, neurotoxic vestibulopathy,pediatric vestibular disorder, Cogan syndrome, vestibular hyperacusis,vertebrobasilar insufficiency.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating non-lesional vestibulardiseases, disorders or conditions, including without limitation, benignpositional paroxysmal vertigo, motion sickness and mal de debarquement.

Methods to determine if a disease, disorder or condition is a lesionalvestibular disease, disorder or condition, i.e., if the functionality ofthe vestibule is impaired, include MRI for identifying large lesions orindirect assessment methods allowing the evaluation of the loss offunctionality of the vestibule. These methods are generally conducted atENT clinic/hospitals and include the videonystagmography (VNG), andassessment of the vestibulo-ocular reflex (VOR) using caloric orrotational tests. The function of the vestibulo-ocular reflex (VOR) isto stabilize the visual image on the retina during displacement.Measurement of this VOR provides convenient method to investigate thefunctionality of the vestibular system. Basically, the paradigm in basedon monitoring eyes movements by infrared light projection technique(Sergi et al., “Cisplatin ototoxicity in the guinea pig: vestibular andcochlear damage”. Hear Res. 2003 August; 182(1-2):56-64). Patients aresinusoidally oscillated in the dark around their vertical andlongitudinal axes in order to evoke horizontal and vertical eyeresponses. Any functional impairment of the vestibule is associated withalterations in the gain of the evoked VNG. Besides VOR and VNG,posturography methods are used to detect postural deviations of the bodythat are also related to impairments of the vestibule. Morphofunctionalinvestigations such as functional imaging (MRI or CAT (computerizedaxial tomography) and derivates) can be used to detect profound lesionswithin the vestibular endorgans. Specifically adapted VNG, VOR andpostural testing are used in animal models of vestibular deficits toevaluate the amplitude of the insults or lesions in the vestibule.Histological studies are also possible using conventional light orelectron microscopy on fixed tissue (vestibular ganglia and vestibularendorgans). Such investigations are mostly done in rodents.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating inner ear diseases,disorders or conditions, including without limitation, ototoxicity(including, without limitation, compound-induced ototoxicity), infectionof the inner ear, trauma (including, without limitation, trauma of theinner ear, blunt or blast-induced head trauma leading to inner eardamage), tumors of the inner ear, inner ear inflammation, inner ear celldegeneration or age-induced inner ear cell degeneration, autoimmune eardisorder, cochlear disorder, vestibular disorder, Meniere's disease(including, without limitation, spell of Meniere's disease, chronicMeniere disease), neuritis (including, without limitation, vestibularneuritis, vestibular neuronitis, labyrinthitis), acute unilateralvestibulopathy, vestibular migraine, migrainous vertigo, otitis media(including, without limitation, chronic otitis media, acute otitismedia, serous otitis media (otitis media with effusion), acute andchronic suppurative otitis media), mastoiditis (including, withoutlimitation, acute and chronic mastoiditis), perforation of the eardrum(tympanic membrane), otosclerosis, superior semicircular canaldehiscence syndrome, otalgia, tinnitus, myoclonus, and mal dedebarquement.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating middle ear diseases,disorders or conditions, including without limitation, hearing loss,tinnitus, infection of the middle ear, Meniere's disease (including,without limitation, spell of Meniere's disease, chronic Menieredisease), otitis media (including, without limitation, chronic otitismedia, acute otitis media, serous otitis media (otitis media witheffusion), acute and chronic suppurative otitis media), mastoiditis(including, without limitation, acute and chronic mastoiditis), adenoidhypertrophy, neoplasia, intratubal obstruction, middle ear obstruction,perforation of the eardrum (tympanic membrane), cholesteatoma(congenital, primary acquired, secondary acquired), CANVAS syndrome,tympanosclerosis, trauma (including, without limitation, trauma of themiddle ear, temporal bone fractures, barotrauma, head trauma,blast-induced trauma), benign and malignant tumors (including, withoutlimitation, tumors of the middle ear, glomus tumors, auditory nervetumor (i.e., acoustic neuroma, acoustic neurinoma, vestibularschwannoma, eighth nerve tumor), malignant neoplasia including squamouscell carcinoma), and otalgia.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating external ear diseases,disorders or conditions, including without limitation, infection of theexternal ear, allergy of the external ear, trauma of the external ear,cysts, tumors, otitis externa (“swimmer's ear”), acute otitis externa,chronic otitis externa, suppurative otitis externa, necrotizing externalotitis, otomycosis, perichondritis, bullous myringitis, granularmyringitis, herpes zoster oticus (Ramsey Hunt syndrome), contactdermatitis, ear eczema, lacerations of the external canal, presence offoreign bodies, pilar (sebaceous) cysts, epidermal cysts, benign lesionsincluding exostosis and malignant lesions including basal cellepithelioma, squamous cell carcinoma and otalgia.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is for treating inflammation of the earinclude without limitation, inner ear inflammation, autoimmune eardisorder, inflammation of the middle ear, otitis media (including,without limitation, chronic otitis media, acute otitis media, serousotitis media (otitis media with effusion), acute and chronic suppurativeotitis media), Muckle-Wells Syndrome, suppurative labyrinthitis,meningitis, mumps or measles.

In one embodiment, the subject is a mammal. In one embodiment, thesubject is a pet, such as a dog, a cat, a guinea pig, a hamster, a rat,a mouse, a ferret, a rabbit, a bird or an amphibian. In one embodiment,the subject is a human.

In one embodiment, the subject is a male. In one embodiment, the subjectis a female.

In one embodiment, the subject is an adult, i.e., a subject aged 18 ormore (in human years). In one embodiment, the subject is a teenager,i.e., a subject aged from 12 to 18 (in human years), such as a subjectaged 12, 13, 14, 15, 16, 17 (in human years). In one embodiment, thesubject is a child, i.e., a subject aged below 12 (in human years), suchas a subject aged 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 (in human years).

In one embodiment, the subject is an adult weighting from about 40 toabout 100 kg or more. In one embodiment, the subject in a teenagerweighting from about 20 to about 80 kg or more. In one embodiment, thesubject is a child weighting from about 1 to about 40 kg or more.

In one embodiment, the subject is awaiting the receipt of, or isreceiving medical care or was/is/will be the object of a medicalprocedure, or is monitored for the development of an ear disease,disorder or condition.

In one embodiment, the subject is/was diagnosed with cancer.

Examples of cancers include, but are not limited to, neuroblastoma,hepatoblastoma, medulloblastoma, osteosarcoma, malignant germ celltumour, nasopharyngeal carcinoma, lung cancer, colon cancer, breastcancer, ovarian cancer, prostate cancer, testicular cancer, stomachcancer, kidney cancer, liver cancer, bladder cancer, esophageal cancer,brain cancer, head and neck cancer, bone cancer, and leukemia.

In one embodiment, the subject is awaiting the receipt of, or isreceiving chemotherapy.

Chemotherapy refers to course of treatment wherein a chemotherapeuticagent is administered to an individual diagnosed with a cancer. Achemotherapeutic agent includes agents such as drugs which canadvantageously be administered to an individual with cancer, to treatsaid cancer. Examples of chemotherapy include, but are not limited to,platinum-based chemotherapy and non-platinum-based chemotherapy.

Examples of platinum-based chemotherapy include, but are not limited to,cisplatin, carboplatin, oxaliplatin, satraplatin, picoplatin,tetraplatin, transplatin, nedaplatin, ormaplatin, PtCl2[R,RDACH],pyriplatin, ZD0473, BBR3464 and Pt-1C3.

Examples of non-platinum-based chemotherapy include, but are not limitedto, nucleoside analogues, antifolates, topoisomerase I inhibitors,anthracyclines, podophyllotoxins, taxanes, vinca alkaloids, alkylatingagents, monoclonal antibodies, tyrosine kinase inhibitor, mTORinhibitors, retinoids, immunomodulatory agents, histone deacetylaseinhibitors and the like.

In one embodiment, the subject is/was diagnosed with cancer and isawaiting the receipt of, or is receiving platinum-based chemotherapy,preferably cisplatin, carboplatin, oxaliplatin or a combination thereof.

In one embodiment, the subject is awaiting receipt of, or is receivingaminoglycosides (such as, but not limited to, neomycin, gentamycin,kanamycin, amikacin and tobramycin), loop diuretics (such as, withoutlimitation, furosemide, torsemide and bumetanide), macrolides (such as,without limitation, erythromycin and azithromycin), glycopeptides (suchas, without limitation, vancomycin) antimetabolites (such as, withoutlimitation, methotrexate), antimalarials (such as, without limitation,quinine and analogs thereof), salicylates and analogs thereof (such as,without limitation, aspirin), radiation treatment, antibiotic agents,excitatory neurotoxins, immunosuppressive agents, β-blockers,vasodilator agents, anti-retroviral drugs (such as, without limitation,abacavir, AZT, delavirdine, didenosine, efavirenz, emtricitabine,indinavir, lamivudine, nefinavir, nevirapine, tenofovir, ritonavir,stavudine and zalcitabine), cardiovascular agents (e.g. ACE inhibitors),antiplatelet agents, radiocontrast media, immunoglobulins, mannitol,NSAIDs, heavy metals, lithium salts, catechol-O-methyltransferaseinhibitors, antiepileptic drugs, proton pump inhibitors,antidepressants, antihistamines (e.g. diphenhydramine, doxylamine),antibacterial sulfonamides, benzodiazepines, pesticides, neurotoxicagents (e.g. glutamate, Bicuculline, Botulinum toxin) or a combinationthereof.

In one embodiment, the subject uses or is exposed or is at risk of beingexposed to, alcohol, heavy metals, drugs (e.g. cocaine, heroin,ketamine) or a combination thereof.

In one embodiment, the compound of Formula (I) or a pharmaceuticallyacceptable salt and/or solvate thereof, preferably (+)-azasetron or apharmaceutically acceptable salt and/or solvate thereof, preferably(+)-azasetron besylate, (+)-azasetron hydrochloride or (+)-azasetronmalate; or the composition comprising thereof, pharmaceuticalcomposition comprising thereof or medicament comprising thereof, is tobe administered at a dose determined by the skilled artisan andpersonally adapted to each subject.

In one embodiment of the invention, the compound of Formula (I) or apharmaceutically acceptable salt and/or solvate thereof, preferably(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof, preferably (+)-azasetron besylate, (+)-azasetron hydrochlorideor (+)-azasetron malate; or the composition comprising thereof,pharmaceutical composition comprising thereof or medicament comprisingthereof is to be administered at a therapeutically effective amount.

It will be understood that the total daily usage of the compound ofFormula (I) or a pharmaceutically acceptable salt and/or solvatethereof, preferably (+)-azasetron or a pharmaceutically acceptable saltand/or solvate thereof, preferably (+)-azasetron besylate, (+)-azasetronhydrochloride or (+)-azasetron malate; or of the composition comprisingthereof, pharmaceutical composition comprising thereof or medicamentcomprising thereof will be decided by the attending physician within thescope of sound medical judgment. The specific therapeutically effectivedose level for any particular patient will depend upon a variety offactors including the disorder being treated and the severity of thedisorder; activity of the specific compound employed; the specificcomposition employed, the age, body weight, general health, sex and dietof the patient; the time of administration, route of administration, andrate of excretion of the specific compound employed; the duration of thetreatment; drugs used in combination or coincidental with the specificcomposition employed; and like factors well known in the medical arts.For example, it is well within the skill of the art to start doses ofthe compound at levels lower than those required to achieve the desiredtherapeutic effect and to gradually increase the dosage until thedesired effect is achieved. However, the daily dosage of the compound ofFormula (I) or a pharmaceutically acceptable salt and/or solvatethereof, preferably (+)-azasetron or a pharmaceutically acceptable saltand/or solvate thereof, preferably (+)-azasetron besylate, (+)-azasetronhydrochloride or (+)-azasetron malate may be varied over a wide rangefrom about 0.1 to about 10000 mg per adult per day, preferably 0.1 toabout 2000, more preferably from about 0.1 to about 500 mg per adult perday, more preferably from about 0.1 to about 100 mg per adult per day.

Preferably, the compositions contain 0.1, 0.5, 1, 10, 20, 50, 100, 250,500, 1000 or 2,000 mg of the compound of Formula (I) or apharmaceutically acceptable salt and/or solvate thereof, preferably(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof, preferably (+)-azasetron besylate, (+)-azasetron hydrochlorideor (+)-azasetron malate, for the symptomatic adjustment of the dosage tothe patient to be treated. In one embodiment, quantities of Formula (I)compound, preferably of (+)-azasetron, are expressed as free-baseequivalent. Therefore, in one embodiment, the compositions contain 1, 5,10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95or 100 mg of the free-base compound of Formula (I), preferably offree-base (+)-azasetron. In one embodiment, quantities of Formula (I)compound, preferably of (+)-azasetron, are expressed as salt and/orsolvate equivalent. Therefore, in one embodiment, the compositionscontain 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75,80, 85, 90, 95, 100, 105, 110, 115, 120 or 125 mg of a pharmaceuticallyacceptable salt and/or solvate of the compound of Formula (I),preferably of a pharmaceutically acceptable salt and/or solvate of(+)-azasetron, preferably (+)-azasetron besylate, (+)-azasetronhydrochloride or (+)-azasetron malate.

A medicament may typically contain from about 0.1 to about 10000 mg ofthe compound of Formula (I) or a pharmaceutically acceptable salt and/orsolvate thereof, preferably (+)-azasetron or a pharmaceuticallyacceptable salt and/or solvate thereof, preferably (+)-azasetronbesylate, (+)-azasetron hydrochloride or (+)-azasetron malate,preferably from about 0.1 to about 2000 mg, more preferably from about0.1 to about 500 mg, more preferably from about 0.1 to about 100 mg ofthe compound of Formula (I) or a pharmaceutically acceptable salt and/orsolvate thereof, preferably (+)-azasetron or a pharmaceuticallyacceptable salt and/or solvate thereof, preferably (+)-azasetronbesylate, (+)-azasetron hydrochloride or (+)-azasetron malate. In oneembodiment, quantities of Formula (I) compound, preferably of(+)-azasetron, are expressed as free-base equivalent. Therefore, in oneembodiment, the medicament contains 1, 5, 10, 15, 20, 25, 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100 mg of the free-basecompound of Formula (I), preferably of free-base (+)-azasetron. In oneembodiment, quantities of Formula (I) compound, preferably of(+)-azasetron, are expressed as salt and/or solvate equivalent.Therefore, in one embodiment, the medicament contains 1, 5, 10, 15, 20,25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105,110, 115, 120 or 125 mg of a pharmaceutically acceptable salt and/orsolvate of the compound of Formula (I), preferably of a pharmaceuticallyacceptable salt and/or solvate of (+)-azasetron, preferably(+)-azasetron besylate, (+)-azasetron hydrochloride or (+)-azasetronmalate.

An effective amount of the free-base compound of Formula (I) or apharmaceutically acceptable salt and/or solvate thereof, preferablyfree-base (+)-azasetron or a pharmaceutically acceptable salt and/orsolvate thereof, preferably (+)-azasetron besylate, (+)-azasetronhydrochloride or (+)-azasetron malate may ordinarily be supplied at adosage level from about 0.01 mg/kg to about 100 mg/kg of body weight perday, preferably from about 0.02 mg/kg to 20 mg/kg of body weight perday, more preferably from about 0.05 mg/kg to 5 mg/kg of body weight perday, more preferably from about 0.1 mg/kg to 2 mg/kg of body weight perday, more preferably at about 0.5 mg/kg of body weight per day. In oneembodiment, effective amounts of Formula (I) compound, preferably of(+)-azasetron, are expressed as free-base equivalent. In one embodiment,quantities of Formula (I) compound, preferably of (+)-azasetron, areexpressed as salt and/or solvate equivalent.

In one embodiment, the dosage of the free-base compound of Formula (I),preferably free-base (+)-azasetron ranges from about 0.1 to about 500 mgof free-base equivalent per adult per day, preferably from about 1 toabout 200 mg, more preferably from about 10 to about 100 mg, morepreferably from about 30 to about 60 mg of free-base equivalent peradult per day. In one embodiment, the dosage of a pharmaceuticallyacceptable salt and/or solvate of the compound of Formula (I),preferably a pharmaceutically acceptable salt and/or solvate of(+)-azasetron, preferably (+)-azasetron besylate, (+)-azasetronhydrochloride or (+)-azasetron malate, ranges from about 0.1 to about500 mg of salt and/or solvate equivalent per adult per day, preferablyfrom about 1 to about 200 mg, more preferably from about 10 to about 100mg, more preferably from about 43 to about 87 mg of salt and/or solvateequivalent per adult per day.

In one embodiment, the subject is a preterm newborn, a term newborn, achild and/or a teenager. In one embodiment, the subject is/was diagnosedwith cancer. In one embodiment, the subject is awaiting receipt of or isreceiving chemotherapy, preferably platinum-based chemotherapy, morepreferably cisplatin-based chemotherapy. Then, in one embodiment, aneffective amount of the compound of Formula (I) or a pharmaceuticallyacceptable salt and/or solvate thereof, preferably (+)-azasetron or apharmaceutically acceptable salt and/or solvate thereof, preferably(+)-azasetron besylate, (+)-azasetron hydrochloride or (+)-azasetronmalate may ordinarily be supplied at a dosage level from about 0.1 mg/kgto about 2 mg/kg, preferably from about 0.2 mg/kg to about 1 mg/kg, morepreferably about 0.5 mg/kg. In one embodiment, the maximum daily dosageof the free-base compound of Formula (I), preferably free-base(+)-azasetron ranges from about 0.1 to about 500 mg of free-baseequivalent per day, preferably from about 1 to about 200 mg, morepreferably from about 10 to about 100 mg, more preferably from about 30to about 60 mg of free-base equivalent per day. In one embodiment, themaximum daily dosage of a pharmaceutically acceptable salt and/orsolvate of the compound of Formula (I), preferably a pharmaceuticallyacceptable salt and/or solvate of (+)-azasetron, preferably(+)-azasetron besylate, (+)-azasetron hydrochloride or (+)-azasetronmalate ranges from about 0.1 to about 500 mg of salt and/or solvateequivalent per day, preferably from about 1 to about 200 mg, morepreferably from about 10 to about 100 mg, more preferably from about 43to about 87 mg of salt and/or solvate equivalent per day.

In one embodiment, a therapeutically effective amount of the compound,composition, pharmaceutical composition or medicament of the inventionis administered at least once a day, twice a day, or at least threetimes a day.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is administered twice a day, at a dosageranging from about 10 mg to about 50 mg each dose, preferably from about20 mg to about 40 mg each dose, more preferably at about 30 mg offree-base equivalent each dose. In one embodiment, each dose maycorrespond to one or more (in particular 2, 3, 4 or 5) unit forms, suchas, for example, one or more oral unit form.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is administered twice a day, at a totaldaily dosage ranging from about 20 mg to about 100 mg, preferably fromabout 40 mg to about 80 mg, more preferably at about 60 mg of free-baseequivalent. In one embodiment, each dose may correspond to one or more(in particular 2, 3, 4 or 5) unit forms, such as, for example, one ormore oral unit form.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is administered once a day, at a dosageranging from about 20 mg to about 100 mg, preferably from about 40 mg toabout 80 mg, more preferably at about 60 mg of free-base equivalent.

In another embodiment, a therapeutically effective amount of thecompound, composition, pharmaceutical composition, or medicament of theinvention is administered every two, three, four, five, or six days.

In another embodiment, a therapeutically effective amount of thecompound, composition, pharmaceutical composition, or medicament of theinvention is administered twice a week, once a week, every two weeks, oronce a month.

In another embodiment, a therapeutically effective amount of thecompound, composition, pharmaceutical composition or medicament of theinvention is administered for a period of time decided by the attendingphysician within the scope of sound medical judgment or for the rest ofthe life of the subject.

In another embodiment, a therapeutically effective amount of thecompound, composition, pharmaceutical composition or medicament of theinvention is administered once, twice, three times, four times, fivetimes, six times, seven times, eight times, nine times, ten times,eleven times, twelve times, thirteen times, fourteen times, fifteentimes, sixteen times, seventeen times, eighteen times, nineteen times,twenty times, twenty-one times, twenty-two times, twenty-three times,twenty-four times, twenty-five times, twenty-six times, twenty-seventimes, twenty-eight times, twenty-nine times, thirty times or more.

In another embodiment, a therapeutically effective amount of thecompound, composition, pharmaceutical composition or medicament of theinvention is administered for a week, 2 weeks, 3 weeks, a month, twomonth, three month, or for the rest of the life of the subject.

In another embodiment, a therapeutically effective amount of thecompound, composition, pharmaceutical composition or medicament of theinvention is administered every month for a period at least 2; 3; 4; 5;6 months or for the rest of the life of the subject.

In one embodiment, when the subject is/was diagnosed with sensorineuralhearing loss, preferably with sudden sensorineural hearing loss, atherapeutically effective amount of the compound, composition,pharmaceutical composition, or medicament of the invention isadministered for a period at least 2, 3, 4, 5 weeks or more, preferablyfor a period of at least 3, 4 or 5 weeks, more preferably for a periodof 4 weeks after the appearance of symptoms or after the initial stagesof case management. In one embodiment, when the subject is/was diagnosedwith sensorineural hearing loss, preferably with sudden sensorineuralhearing loss, a therapeutically effective amount of the compound,composition, pharmaceutical composition, or medicament of the inventionis administered for a period at least 21, 22, 23, 24, 25, 26, 27, 28,29, 30, 31, 32, 33, 34, 35 days or more, preferably for a period of atleast 25, 26, 27, 28, 29, 30 or 31 days, more preferably for a period of28 days after the appearance of symptoms or after the initial stages ofcase management.

In one embodiment, the subject receives the first administration of thecompound, composition, pharmaceutical composition, or medicament of theinvention at most one week, preferably at most 6, 5, 4, 3, 2 or 1 dayafter the appearance of symptoms, including, without limitation, hearingloss or tinnitus.

In one embodiment, when the subject is/was diagnosed with cancer, atherapeutically effective amount of the compound, composition,pharmaceutical composition, or medicament of the invention isadministered before, during and/or after chemotherapy, preferablyplatinum-based chemotherapy, more preferably cisplatin-, carboplatin- oroxaliplatin-based chemotherapy or a combination thereof.

In one embodiment, a therapeutically effective amount of the compound,composition, pharmaceutical composition, or medicament of the inventionis administered before platinum-based chemotherapy, preferablycisplatin-, carboplatin- or oxaliplatin-based chemotherapy or acombination thereof, such as for 1 day, 2 days, 3 days, 4 days, 5 days,6 days, 7 days or more before.

In one embodiment, a therapeutically effective amount of the compound,composition, pharmaceutical composition, or medicament of the inventionis administered after platinum-based chemotherapy, preferablycisplatin-, carboplatin- or oxaliplatin-based chemotherapy or acombination thereof, such as for 1 day, 2 days, 3 days, 4 days, 5 days,6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days or moreafter.

In one embodiment, a therapeutically effective amount of the compound,composition, pharmaceutical composition, or medicament of the inventionis administered immediately before and/or immediately afterplatinum-based chemotherapy, preferably cisplatin-, carboplatin- oroxaliplatin-based chemotherapy or a combination thereof.

In one embodiment, a therapeutically effective amount of the compound,composition, pharmaceutical composition, or medicament of the inventionis administered for one day, two days, three days, four days or morebefore and for 1 day, two days, three days, four days, five days, sixdays, a week, two weeks, three weeks or more after platinum-basedchemotherapy, preferably cisplatin-, carboplatin- or oxaliplatin-basedchemotherapy or a combination thereof. In one embodiment, atherapeutically effective amount of the compound, composition,pharmaceutical composition, or medicament of the invention isadministered for at least one day before and for at least 14 days afterplatinum-based chemotherapy. In one embodiment, a therapeuticallyeffective amount of the compound, composition, pharmaceuticalcomposition, or medicament of the invention is administered for one daybefore and for 14 days after platinum-based chemotherapy.

In one embodiment, a therapeutically effective amount of the compound,composition, pharmaceutical composition, or medicament of the inventionis administered at each cycle of platinum-based chemotherapy, such as,for example, at each of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more cycles ofplatinum-based chemotherapy, preferably at each of 6 cycles ofplatinum-based chemotherapy.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is to be administered systemically orlocally.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is to be administered by injection,orally, topically, nasally, by inhalation, buccally, rectally,intratracheally, transmucosally, transtympanically, by percutaneousadministration, intramuscularly or by parenteral administration.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is to be administered by injection,preferably is to be systemically injected. Examples of formulationsadapted to systemic injections include, but are not limited to: liquidsolutions or suspensions, solid forms suitable for solution in, orsuspension in, liquid prior to injection. Examples of systemicinjections include, but are not limited to, intravenous, subcutaneous,intramuscular, intradermal, intravitreal, and intraperitoneal injection,or perfusion. In another embodiment, when injected, the compound,composition, pharmaceutical composition or medicament of the inventionis sterile. Methods for obtaining a sterile pharmaceutical compositioninclude, without limitation, sterile filtration, terminal sterilization(dry heat, radiation, moist heat, gases, gamma radiation) orsterilization via aseptic processing.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is to be systemically administered,preferably is to be orally administered. Examples of formulationsadapted to oral administration include, but are not limited to: solidforms, liquid forms and gels. Examples of solid forms adapted to oraladministration include, but are not limited to, pill, tablet, capsule,soft gelatine capsule, hard gelatine capsule, caplet, compressed tablet,cachet, wafer, sugar-coated pill, sugar coated tablet, or dispersingand/or disintegrating tablet, powder, solid forms suitable for solutionin, or suspension in, liquid prior to oral administration andeffervescent tablet. Examples of liquid forms adapted to oraladministration include, but are not limited to, solutions, suspensions,drinkable solutions, elixirs, sealed phial, potion, drench, syrup andliquor.

In another embodiment, the compound, composition, pharmaceuticalcomposition or medicament of the invention is to be topicallyadministered. Examples of formulations adapted to topical administrationinclude, but are not limited to, sticks, waxes, creams, lotions,ointments, balms, gels, masks, leave-on washes and/or the like.

In one embodiment, the compound, composition, pharmaceutical compositionor medicament of the invention is to be administered directly in theear, in particular, in the inner ear, in the middle ear, in the externalear, in the cochlea, or in the vestibule by transtympanic orintratympanic administration. This administration route may be preferredfor introducing a direct and long term effect on the ear. Saidadministration can be accomplished topically or by injection. Deliverytechniques for such administration may include the use of devices ordrug carriers to transport and/or deliver the active principle to theear, where it diffuses into the ear, is actively infused or is injected.Examples of formulations adapted to such administration include, but arenot limited to, otowicks, round window catheters, various types of gels,foams, fibrins, emulsions, solutions, patches or other drug carriers,which are placed in the ear, and loaded with the composition of theinvention for sustained release. It may also include devices that areinserted into the cochlear duct or any other part of the cochlea.

The diffusion of the composition across middle-inner ear interfacetissue structures, in particular the round window membrane, depends on avariety of factors, such as molecular weight, concentration,liposolubility, electrical charge, and thickness of the membrane(Goycoolea M. and Lundman L., Microscopy Research and Technique 36:201-211 (1997).

In one embodiment, the compound described here above is to beadministered in a immediate-release form.

In one embodiment, the compound described here above is to beadministered in a sustained-release form. In another embodiment, thecomposition, the pharmaceutical composition or the medicament of theinvention comprises a delivery system that controls the release of theat least one compound of Formula I, preferably of (+)-azasetron or apharmaceutically acceptable salt and/or solvate thereof, more preferablyof (+)-azasetron besylate, (+)-azasetron hydrochloride, (+)-azasetronmalate or a mixture thereof.

In another embodiment, the composition, pharmaceutical composition ormedicament of the invention comprises sustained-release drug deliveryagents, such as biodegradable polymers. As used herein, asustained-release drug delivery agent is a composition, e.g., apolymeric matrix, which provides a reservoir or vehicle for release of atherapeutic agent over an extended time in a subject, e.g., in asubject's ear canal.

In some embodiments, a sustained-release drug delivery agent is amaterial, such as, for example, a polyelectrolyte or thermo-responsivepolymer, that undergoes a viscosity increase after being administered toa subject, e.g., administered into a subject's ear canal.

It should be appreciated that the sustained-release drug delivery agentsinclude a variety of materials, including, without limitation, polymericmaterials that form in response to temperature change (e.g.,poloxamers), polyelectrolyte complexing (e.g., chitosan/chondroitinsulfate), polymer cross-linking (both physical and chemical, e.g., withrheological synergism or hyaluronic acid derivatives, respectively), orsensitivity to photo or electromagnetic waves (e.g., UV or microwaves),solvent exchange, or pH. In certain embodiments, the sustained-releasedrug delivery agent is a hydrophilic material.

In some embodiments, the sustained-release drug delivery agent is amatrix-forming agent. Matrix-forming agents are generally liquid atambient conditions, however, once administered to a subject, the matrixforming agent gels (i.e., becomes more viscous). In various aspects, forexample, the matrix-forming agent changes viscosity once administeredinto a patient's ear canal forming in situ a reservoir in contact withor nearby the tympanic membrane. A reservoir in contact with thetympanic membrane maximizes exposure and concentration of thetherapeutic agent at the surface of the tympanic membrane, thusincreasing flux of the agent across the tympanic membrane and into themiddle and/or inner ear. Exemplary matrix-forming agents include, butare not limited to, polyelectrolyte complexes (e.g.,chitosan-chondroitin complexes), thermo-responsive gelling agents (e.g.,poloxamers), pre-polymers, alginates, un-crosslinked polymers, andmonomers.

In the compositions of the present invention, the active principle,alone or in combination with another active principle, can beadministered in a unit administration form, as a mixture withconventional pharmaceutical supports, to animals and human beings.

Suitable unit administration forms comprise oral-route forms such astablets, gel capsules, powders, granules and oral suspensions orsolutions, sublingual and buccal administration forms, gingival ormucosal or mucoadhesive formulations, aerosols, sprays, transtympanical,implants, subcutaneous, transdermal, topical, intraperitoneal,intramuscular, intravenous, subdermal, transdermal, intrathecal andintranasal administration forms and rectal administration forms.

In one embodiment, the active principle, alone or in combination withanother active principle, can be administered in an oral-route form(immediate or sustained-released), preferably as tablets.

In one embodiment, a tablet for oral-route administration comprises orconsists of or consists essentially of at least one compound of FormulaI or a pharmaceutically acceptable salt and/or solvate thereof,preferably (+)-azasetron or a pharmaceutically acceptable salt and/orsolvate thereof, more preferably (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof.

In one embodiment, the compound of Formula I or a pharmaceuticallyacceptable salt and/or solvate thereof, preferably (+)-azasetron or apharmaceutically acceptable salt and/or solvate thereof, more preferably(+)-azasetron besylate, (+)-azasetron hydrochloride, (+)-azasetronmalate or a mixture thereof may be formulated within a tablet fororal-route administration comprising or consisting of about 0.1 to 10000milligrams, preferably from about 0.1 to 1000 milligrams, morepreferably from about 0.1 to 100 milligrams, even more preferably fromabout 1 to 100 milligrams per dose or so.

In one embodiment, a tablet for oral-route administration comprises orconsists of at least about 0.01 mg, 0.05 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 20mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150mg, 175 mg, 200 mg or more of compound of Formula I or apharmaceutically acceptable salt and/or solvate thereof, preferably(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof, more preferably (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof. In oneembodiment, quantities of Formula (I) compound, preferably of(+)-azasetron, are expressed as free-base equivalent. Therefore, in oneembodiment, a tablet for oral-route administration comprises or consistsof at least about 0.01 mg, 0.05 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg,7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 20 mg, 25mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg,175 mg, 200 mg or more of the free-base compound of Formula (I),preferably of free-base (+)-azasetron. In one embodiment, quantities ofFormula (I) compound, preferably of (+)-azasetron, are expressed as saltand/or solvate equivalent. Therefore, in one embodiment, a tablet fororal-route administration comprises or consists of at least about 0.01mg, 0.05 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8mg, 0.9 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg,11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg or more of apharmaceutically acceptable salt and/or solvate of the compound ofFormula (I), preferably of a pharmaceutically acceptable salt and/orsolvate of (+)-azasetron, preferably (+)-azasetron besylate,(+)-azasetron hydrochloride or (+)-azasetron malate.

In one embodiment, a tablet for oral-route administration may comprisesone or more disintegrants, one or more binders, one or more lubricantsand/or one or more coatings.

In one embodiment, a tablet for oral-route administration comprises atleast one disintegrant. Examples of disintegrants include, but are notlimited to, microcrystalline cellulose (MCC), carboxymethylstarch-sodium(such as Pirimojel and Explotab), carboxymethylcellulose calcium(CMC-Ca), carboxymethylcellulose sodium (CMC-Na), crosslinked CMC (suchas Ac-Di-Sol), crosslinked PVP (such as Crospovidone, Polyplasdone orKollidon XL), alginic acid, sodium alginate, guar gum.

In one embodiment, a tablet for oral-route administration comprises atleast one binder. Examples of binders include, but are not limited to,starch, polyvinylpyrrolidone (PVP), hydroxypropylmethylcellulose (HPMC)and polyethylene glycols (PEG).

In one embodiment, a tablet for oral-route administration comprises atleast one lubricant. Examples of lubricant include, but are not limitedto, magnesium stearate, aluminum stearate, calcium stearate,polyethylene glycol (PEG) 4000 to 8000, talc, hydrogenated castor oil,stearic acid and salts thereof, glycerol esters, Na-stearylfumarate,hydrogenated cotton seed oil and the like.

In one embodiment, a tablet for oral-route administration comprises atleast one coating. Examples of coatings include, but are not limited to,hydroxypropylmethylcellulose (HPMC), polyethylene glycol (PEG),polysorbate 80, polyvinylpyrrolidone (PVP), PVP-vinyl acetate copolymer(PVP-VA), polyvinyl alcohol (PVA), and sugar. Further coatings comprisepigments, dies, lakes, titanium dioxide, iron oxides, anti-tackingagents (such as talk and softeners like PEG 3350, 4000, 6000, 8000) orothers.

In one embodiment, a tablet for oral-route administration is animmediate release tablet. In one embodiment, a tablet for oral-routeadministration is an orodispersible tablet. In one embodiment, a tabletfor oral-route administration is a sustained-release tablet.

The composition, pharmaceutical composition or medicament of theinvention may contain vehicles which are pharmaceutically acceptable fora formulation capable of being injected. These may be in particularisotonic, sterile, saline solutions (monosodium or disodium phosphate,sodium, potassium, calcium or magnesium chloride and the like ormixtures of such salts), or dry, especially freeze-dried compositions orlyophilisates which upon addition, depending on the case, of sterilizedwater or physiological saline, permit the constitution of injectablesolutions.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions; formulations including sesame oil,peanut oil or aqueous propylene glycol; and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases, the form must be sterile and must be fluid tothe extent that easy administration via syringe or needle is obtained.It must be stable under the conditions of manufacture and storage andmust be preserved against the contaminating action of microorganisms,such as bacteria and fungi.

Solutions comprising at least one compound of the invention as free-baseor pharmacologically acceptable salts can be prepared in water suitablymixed with a surfactant, such as hydroxypropylcellulose. Dispersions canalso be prepared in glycerol, liquid polyethylene glycols, and mixturesthereof and in oils. Under ordinary conditions of storage and use, thesepreparations may contain a preservative to prevent the growth ofmicroorganisms.

The compounds of the invention can be formulated into a composition in aneutral or salt form. Pharmaceutically acceptable salts include the acidaddition salts which are formed with inorganic acids such as, forexample, hydrochloric or phosphoric acids, or such organic acids asacetic, benzenesulfonic, malic, oxalic, tartaric, mandelic, and thelike.

The carrier can also be a solvent or dispersion medium containing, forexample, water, ethanol, polyol (for example, glycerol, propyleneglycol, and liquid polyethylene glycol, and the like), suitable mixturesthereof, and vegetables oils. The proper fluidity can be maintained, forexample, by the use of a coating, such as lecithin, by the maintenanceof the required particle size in the case of dispersion and by the useof surfactants. The prevention of the action of microorganisms can bebrought about by various antibacterial and antifungal agents, forexample, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, andthe like. In many cases, it will be preferable to include isotonicagents, for example, sugars or sodium chloride. Prolonged absorption ofthe injectable compositions can be brought about by the use in thecompositions of agents delaying absorption, for example, aluminiummonostearate and gelatin.

Sterile injectable solutions may be prepared by incorporating thecompound of the invention in the required amount in the appropriatesolvent with one or several of the other ingredients enumerated above,as required, followed by sterile filtration or other processingtechnique capable of rendering the solution sterile. Generally,dispersions are prepared by incorporating the various sterilized activeingredients into a sterile vehicle which contains the basic dispersionmedium and the required other ingredients from those enumerated above.In the case of sterile powders for the preparation of sterile injectablesolutions, the preferred methods of preparation are formulation of thebulk solution, followed by sterile filtration and freeze-dryingtechniques which yield a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

Upon formulation, solutions will be administered in a manner compatiblewith the dosage formulation and in such amount as is therapeuticallyeffective. The formulations are easily administered in a variety ofdosage forms, such as the type of injectable solutions described above,but drug release capsules and the like can also be employed.

For parenteral administration in an aqueous solution, for example, thesolution should be suitably buffered if necessary and the liquid diluentfirst rendered isotonic with sufficient saline or glucose. The liquidcompositions may include aqueous solutions with and without organicco-solvents, aqueous or oil suspensions, emulsions with edible oils,liposomes or nano-emulsion as well as similar pharmaceutical vehicles.These particular aqueous solutions are especially suitable fortranstympanic, intravenous, intramuscular, subcutaneous andintraperitoneal administration. Advantageously the viscosity of theformulation for transtympanic solution is between 5000 and 25000 mPas(milli-Pascal second), preferably between 15000 and 20000 mPas, so thata longer period of administration of the active principle into the innerear is achieved. In this connection, sterile aqueous media which can beemployed will be known to those of skill in the art in light of thepresent disclosure. For example, one dosage could be dissolved in 1 mLof isotonic NaCl solution and either added to 1000 mL of hypodermoclysisfluid or injected at the proposed site of infusion. Some variation indosage, pH (preferably between about 5 and 9), and formulation willnecessarily occur depending on the condition of the subject beingtreated, the routes of administration, the compound used.

The person responsible for administration will, in any event, determinethe appropriate dose for the individual subject. The compounds of theinvention may be formulated within a therapeutic mixture to compriseabout 0.1 to 10000 milligrams, preferably from about 0.2 to 1000milligrams, more preferably from about 0.3 to 100 milligrams per dose orso. Multiple doses can also be administered.

Another object of the invention is a method for treating an ear disease,disorder or condition in a subject in need thereof, comprising orconsisting of administering to the subject a therapeutically effectiveamount of at least one compound of Formula I or a pharmaceuticallyacceptable salt and/or solvate thereof.

In one embodiment, the method of the invention is for preventing an eardisease, disorder or condition. In another embodiment, the method of theinvention is for alleviating a symptom of an ear disease, disorder orcondition or for curing an ear disease, disorder or condition.

In one embodiment, the method comprises or consists of administering tothe subject a therapeutically effective amount of free-base(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof (in particular, (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof).

Another object of the invention is a method for treating hearing loss,preferably sensorineural hearing loss in a subject in need thereof,comprising or consisting of administering to the subject atherapeutically effective amount of at least one compound of Formula Ior a pharmaceutically acceptable salt and/or solvate thereof.

In one embodiment, the method of the invention is for preventing hearingloss, preferably sensorineural hearing loss. In another embodiment, themethod of the invention is for alleviating a symptom of hearing loss,preferably of sensorineural hearing loss or for curing hearing loss,preferably sensorineural hearing loss.

In one embodiment, the method comprises or consists of administering tothe subject a therapeutically effective amount of free-base(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof (in particular, (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof).

Another object of the invention is a method for treating ototoxiccompound-induced hearing loss, preferably platinum-induced hearing lossin a subject in need thereof, comprising or consisting of administeringto the subject a therapeutically effective amount of at least onecompound of Formula I or a pharmaceutically acceptable salt and/orsolvate thereof.

In one embodiment, the method of the invention is for preventingototoxic compound-induced hearing loss, preferably platinum-inducedhearing loss. In another embodiment, the method of the invention is foralleviating a symptom of ototoxic compound-induced hearing loss,preferably of platinum-induced hearing loss or for curing ototoxiccompound-induced hearing loss, preferably platinum-induced hearing loss.

In one embodiment, the method comprises or consists of administering tothe subject a therapeutically effective amount of free-base(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof (in particular, (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof).

Another object of the invention is a method for treatingcisplatin-induced hearing loss in a subject in need thereof, comprisingor consisting of administering to the subject a therapeuticallyeffective amount of at least one compound of Formula I or apharmaceutically acceptable salt and/or solvate thereof.

In one embodiment, the method of the invention is for preventingcisplatin-induced hearing loss. In another embodiment, the method of theinvention is for alleviating a symptom of cisplatin-induced hearing lossor for curing cisplatin-induced hearing loss.

In one embodiment, the method comprises or consists of administering tothe subject a therapeutically effective amount of free-base(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof (in particular, (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof).

Another object of the invention is a method for restoring vestibularfunctionality in a subject in need thereof, preferably in a subjectaffected with a lesional vestibular disease, comprising or consisting ofadministering to the subject a therapeutically effective amount of atleast one compound of Formula I or a pharmaceutically acceptable saltand/or solvate thereof.

In one embodiment, the method of the invention is for preventing alesional vestibular disease. In another embodiment, the method of theinvention is for alleviating a symptom of a lesional vestibular diseaseor for curing a lesional vestibular disease.

In one embodiment, the method comprises or consists of administering tothe subject a therapeutically effective amount of free-base(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof (in particular, (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof).

Another object of the invention is a method for restoring hearing and/orbalance in a subject in need thereof, comprising or consisting ofadministering to the subject a therapeutically effective amount of atleast one compound of Formula I or a pharmaceutically acceptable saltand/or solvate thereof.

In one embodiment, the method comprises or consists of administering tothe subject a therapeutically effective amount of free-base(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof (in particular, (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof).

Another object of the invention is a method for reducing loss of haircells in the inner ear of a subject in need thereof, comprising orconsisting of administering to the subject a therapeutically effectiveamount of at least one compound of Formula I or a pharmaceuticallyacceptable salt and/or solvate thereof.

In one embodiment, the method of the invention is for preventing theloss of hair cells in the inner ear. In another embodiment, the methodof the invention is for alleviating a symptom of the loss of hair cellsin the inner ear or for curing the loss of hair cells in the inner ear.

In one embodiment, the method comprises or consists of administering tothe subject a therapeutically effective amount of free-base(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof (in particular, (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof).

Another object of the invention is a method for reducing loss ofsynapses between hair cells and neurons in the inner ear of a subject inneed thereof, comprising or consisting of administering to the subject atherapeutically effective amount of at least one compound of formula Ior a pharmaceutically acceptable salt and/or solvate thereof.

In one embodiment, the method of the invention is for preventing theloss of synapses between hair cells and neurons in the inner ear. Inanother embodiment, the method of the invention is for alleviating asymptom of the loss of synapses between hair cells and neurons in theinner ear or for curing the loss of synapses between hair cells andneurons in the inner ear.

In one embodiment, the method comprises or consists of administering tothe subject a therapeutically effective amount of free-base(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof (in particular, (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof).

Another object of the invention is a method for reducing loss of neuronsin the inner ear of a subject in need thereof, comprising or consistingof administering to the subject a therapeutically effective amount of atleast one compound of Formula I or a pharmaceutically acceptable saltand/or solvate thereof.

In one embodiment, the method of the invention is for preventing theloss of neurons in the inner ear. In another embodiment, the method ofthe invention is for alleviating a symptom of the loss of neurons in theinner ear or for curing the loss of neurons in the inner ear.

In one embodiment, the method comprises or consists of administering tothe subject a therapeutically effective amount of free-base(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof (in particular, (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof).

Another object of the invention is a method for reducing loss ofsupporting cells in the inner ear of a subject in need thereof,comprising or consisting of administering to the subject atherapeutically effective amount of at least one compound of Formula Ior a pharmaceutically acceptable salt and/or solvate thereof.

In one embodiment, the method of the invention is for preventing theloss of supporting cells in the inner ear. In another embodiment, themethod of the invention is for alleviating a symptom of the loss ofsupporting cells in the inner ear or for curing the loss of supportingcells in the inner ear.

In one embodiment, the method comprises or consists of administering tothe subject a therapeutically effective amount of free-base(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof (in particular, (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof).

In one embodiment, the loss of hair cells, synapses, neurons orsupporting cells may be induced by an acoustic trauma, such as, forexample, loud noise exposure, chronic noise exposure, aging, illnesses(including but not limited to high blood pressure and diabetes),ototoxic drugs, head trauma, tumors, blast exposure, autoimmune innerear disease, idiopathic causes, viral or bacterial infections.

Another object of the invention is a method for inhibiting inflammationin the ear (including, without limitation, inflammation in the innerear, middle ear, cochlea and/or vestibule) of a subject in need thereof,comprising or consisting of administering to the subject atherapeutically effective amount of at least one compound of Formula Ior a pharmaceutically acceptable salt and/or solvate thereof.

In one embodiment, the method of the invention is for preventinginflammation in the ear (including, without limitation, inflammation inthe inner ear, middle ear, cochlea and/or vestibule). In anotherembodiment, the method of the invention is for alleviating a symptom ofinflammation in the ear (including, without limitation, inflammation inthe inner ear, middle ear, cochlea and/or vestibule) or for curinginflammation in the ear (including, without limitation, inflammation inthe inner ear, middle ear, cochlea and/or vestibule).

In one embodiment, the method comprises or consists of administering tothe subject a therapeutically effective amount of free-base(+)-azasetron or a pharmaceutically acceptable salt and/or solvatethereof (in particular, (+)-azasetron besylate, (+)-azasetronhydrochloride, (+)-azasetron malate or a mixture thereof).

Preferably, the methods of the invention comprise or consist ofadministering (+)-azasetron besylate, (+)-azasetron hydrochloride or(+)-azasetron malate to the subject.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a set of three graphs comparing the concentration (nM) of theracemic and two enantiomers of azasetron in the perilymph, the innerear, and the peripheral plasma (tail plasma) after oral administration,at termination time-point. Average per time-point subgroups withintreatment groups (+/−SE).

FIG. 2 is a graph comparing the effects of the racemic and twoenantiomers of azasetron on auditory brainstem response (ABR) thresholdshifts in a sensorineural hearing loss (SSNHL) model.

FIG. 3 is a set of two graphs showing the effect of the (+) enantiomerof azasetron at different doses on auditory brainstem response (ABR)threshold shifts and distortion product otoacoustic emission (DPOAE)amplitude loss in a SSNHL model. Statistical analyses were performedusing 2-way analysis of variance (ANOVA) (treatment group, stimulusfrequency) with Holm-Sidak post-test versus vehicle controls. A P valueof <0.05 was statistically significant.

FIG. 4 is a set of two graphs showing the effect of azasetron racematetreatment against acoustic trauma by oral administration.

FIG. 5 is a set of two graphs showing auditory brainstem response (ABR)threshold shifts (A) and distortion product otoacoustic emission (DPOAE)amplitude loss (B) following cisplatin infusion after 14 days of oraltreatment with placebo (n=4) or 26.4 mg/kg (+)-azasetron besylate (n=5).

FIG. 6 is a graph showing the survival of cochlear outer hair cellsfollowing cisplatin infusion after 14 days of oral treatment withplacebo (n=4) or 26.4 mg/kg (+)-azasetron besylate (n=5).

FIG. 7 is a set of two graphs showing auditory brainstem response (ABR)threshold shifts (A) and distortion product otoacoustic emission (DPOAE)amplitude loss (B) following cisplatin infusion after 14 days of oraltreatment with placebo (n=6), 6.6 mg/kg (+)-azasetron besylate (n=7) or13.2 mg/kg (+)-azasetron besylate (n=5).

FIG. 8 is a graph showing the survival of cochlear outer hair cellsfollowing cisplatin infusion after 14 days of oral treatment withplacebo (n=5), 6.6 mg/kg (n=5) or 13.2 mg/kg (n=5) (+)-azasetronbesylate treatment.

FIG. 9 is a set of two graphs showing the effect of (+)-azasetronbesylate (5.6 mg/kg; n=16) versus placebo (n=18) on unilateralvestibular dysfunction induced by intratympanic kainic injection overtime (in hours). A, Spontaneous nystagmus. B, Roll angle deviation.

EXAMPLES

The present invention is further illustrated by the following examples.

Example 1: Pharmacokinetics Study Material and Methods Test AnimalsAssignment to the Study

The 72 male Wistar rats received on 28 Jan. 2016 from Janvier (Janvier,Le Genest-St-Isle) were assigned to the study. They were divided in 3groups of 24 animals each, one group per test article. Each group wasdivided in 7 subgroups according to the 6-termination time-points.

72 Wistar rats were subjected to oral single dose administration of thetest article, racemic azasetron hydrochloride, azasetron hydrochloride(+) or azasetron hydrochloride (−), and sacrificed at differenttime-points to assess the exposure of the test article in the inner ear,perilymph, and plasma.

-   -   Group racemic azasetron: 24 male Wistar, 4 rats per time-point:        10 mg/kg azasetron HCl dissolved in NaCl 0.9%. Administrated as        5 mL/kg p.o.;    -   Group azasetron (+): 24 male Wistar, 4 rats per time-point: 10        mg/kg (+)-azasetron HCl dissolved in NaCl 0.9%. Administrated as        5 mL/kg p.o.;    -   Group azasetron (−): 24 male Wistar, 4 rats per time-point: 10        mg/kg (−)-azasetron HCl dissolved in NaCl 0.9%. Administrated as        5 mL/kg p.o.

There were 6 termination time-points for each test article. Attime-points 15/30/60/120/240/480 minutes after administration:peripheral plasma from the tail was sampled. The animals were sacrificedby i.v. administration of 0.25 mL/rat of a 160 mg/mL pentobarbitalsolution. Following sacrifice, the heart blood and both tympanic bullas(left and right) were sampled.

Sample Preparation for Bio-Analytics Perilymph and Inner Ear Samples

Both tympanic bullas (left and right) were collected, however, only theleft one was used for the perilymph and inner ear analysis. The rightbulla was stored at −20° C. for future needs when required. For animalIDs 1 and 4, the right bulla was used for the sampling of perilymph andinner ear.

The left bulla was dissected fresh. Briefly, the bulla was opened viathe auditory opening to reveal the bone surrounding the cochlea afterwhich the tip of the bone was removed with forceps and a 10 μL pipettetip was inserted to withdraw perilymph.

Upon exposure of the tip of the cochlea, 2 μL of perilymph (fluid) couldbe recovered from the bulla. The perilymph was brought to 10 μL by theaddition of 8 μL water, and then extract via the addition of 30 μL ofacetonitrile. After that, the samples were centrifuged and thesupernatant was transferred to a glass vial appropriate for the HPLC-MSauto-sampler.

Following recovering of the perilymph, the bones of the inner ear andcochlea components were removed with fine forceps and added to a taredEppendorf tube. The collection was weighed and brought to 10 mg with theaddition of water and placed on ice until extraction. 30 μL ofacetonitrile was added and the tubes were incubated in an ultrasoundbath, after which they were centrifuged and the supernatant wastransferred to a glass vial appropriate for the HPLC-MS auto-sampler. Inthe process or removing the bones of the inner ear, they were finelydistributed in the media and broken by forceps. (Due to the very lowvolume of the tissue and its fine structure, separate grinding isconsidered risky vs the potential losses to plastic and media. Thematerials are very fine in nature and the bone particles are small thusproviding only short interfaces to media. The absence of fat and theincubation in solvent is intended to provide sufficient solubility ofactive ingredient).

The acetonitrile used in the sample preparation for bio-analysiscontained the internal standard for the HPLC-MS analysis.

Tail Plasma

30 μL acetonitrile including the internal standard were added to each 10μL plasma sample. After adding the acetonitrile, the samples were mixedby vortex and further centrifuged. The supernatant was transferred to aglass vial appropriate for the HPLC-MS auto-sampler.

Heart Plasma

The plasma samples from the heart blood are stored at −20° C. for futureoptional analysis when needed.

LC-MS/MS Apparatus

The LC system comprised an Agilent 1260 liquid chromatography equippedwith a binary pump and a column oven, together with a PAL HTS-xtauto-sampler. It is linked to the AB SCIEX TRIPLE QUAD™ 4500(triple-quadrupole) instrument with an ESI interface and a switchingvalve integrated (Mass spectrometric analysis). The data acquisition andcontrol system were created using Analyst Software from AppliedBiosystems Inc.

The test article, azasetron hydrochloride, used for the standard curvewas provided by the Sponsor. Water and acetonitrile were HPLC grade. Allother solvents and chemicals were analytical grade or better.

Results Plasma Kinetic and Distribution to Inner Ear of AzasetronFollowing a Single Oral Dose of Racemic Azasetron, (+)-AzasetronHydrochloride and (−)-Azasetron Hydrochloride in Wistar Rats

This study was conducted to compare exposure to azasetron followingadministration of a single oral dose (10 mg/kg) of racemic azasetronhydrochloride, (+)-azasetron hydrochloride and (−)-azasetronhydrochloride. The concentration of the test articles in the inner earsamples were analyzed by HPLC-MS and quantified by comparison with aknown concentration standard curve (Table 1).

TABLE 1 Concentration (ng azasetron free-base/g tissue) of racemicazasetron, (+)-azasetron and (−)-azasetron in the inner ear tissue attermination time-point. Average per time-point subgroups withintreatment groups, and standard error (SE). Average SE Dose Inner earInner ear Treatment (mg/kg) Route Time (ng/g) (ng/g) Racemic azasetron10 p.o. 15 25 13 30 61 13 60 63 3 120 35 9 240 37 2 480 7 2(+)-azasetron 10 p.o. 15 57 29 30 86 50 60 93 34 120 79 21 240 52 8 4806 0 (−)-azasetron 10 p.o. 15 29 15 30 27 16 60 8 1 120 46 11 240 21 6480 3 1

The concentration of the racemic and two enantiomers of azasetronhydrochloride in the perilymph, the inner ear and the peripheral plasmais represented in FIG. 1 and Table 2.

TABLE 2 (AUC) Area under the curve (nM min) of the azasetron free-basepharmacokinetics. AUC nM min Tail Perilymph Inner ear plasma Dose AUC nMAUC nM AUC nM Treatment (mg/kg) Route Time min min min Racemic 10 p.o 151099 712 10078 azasetron 30 1965 2019 15807 60 2729 5319 18556 120 38118394 22450 240 7026 12260 45507 480 9243 14889 64797 Total AUC 2587343592 177194 (+)- 10 p.o 15 1161 1225 5897 azasetron 30 1783 3059 1154860 3102 7643 21012 120 7719 14739 37299 240 13642 22527 66332 480 1350419844 68681 Total AUC 40910 69036 210768 (−)- 10 p.o 15 304 614 1217azasetron 30 613 1189 2037 60 898 1484 3092 120 2179 4631 10183 240 448211560 24941 480 3425 8351 23978 Total AUC 11901 27829 65448

Following administration of a single equivalent oral dose of eachcompound, greater exposure (total AUC) to azasetron was observed in theperilymph, inner ear, and plasma after administration of (+)-azasetronthan that observed after administration of racemic azasetron or(−)-azasetron.

Example 2: Activity of (+)-Azasetron Besylate on Noise-Induced HearingLoss Material and Methods Animals

All experiments were performed using 7-week-old male Wistar rats (CERJ,Le Genest, France) in accordance with the French Ministry of Agricultureregulations and European Community Council Directive no. 86/609/EEC, OJL358. The rats were fed a standard diet ad libitum and maintained on a 12hours' light-dark cycle.

Audiometry

Auditory Brainstem Reponses (ABR) and Distortion Product OtoacousticEmssions (DPOAE) were recording using a RZ6 Auditory Workstation(Tucker-Davis Technologies, Alachua, Fla., USA) with animals deeplyanesthetized using 90 mg/kg ketamine and 10 mg/kg xylazine and placed ona 35° C. recirculating heating pad inside a sound attenuating cubicle(Med Associates Inc., St. Albans, Vt., USA) throughout the experiment.

For ABR recordings, three stainless steel needle electrodes were placedsubdermally over the vertex, the right mastoid and right hind leg ofeach animal. Tone-pips (5 msec duration presented at a rate of 21/s) at8, 16 and 24 kHz were delivered to the right ear using a calibrated MF-1speaker in closed-field configuration (Tucker-Davis Technologies,Alachua, Fla., USA) at attenuating intensity until no reproducibleresponse could be recorded. Close to the ABR threshold, the responses to1000 acoustic stimuli in 5 dB steps were averaged. Responses werelow-pass filtered at 3 kHz.

DPOAEs were recorded and using ER10B+ Low Noise DPOAE microphone(Etymotic Research, Inc., Elk Grove Village, Ill., USA) with acousticstimuli delivered by two calibrated MF-1 speakers in closed-fieldconfiguration (Tucker-Davis Technologies, Alachua, Fla., USA). DPOAEswere recorded at fixed stimulus levels (L1=L2=70 dB SPL), with an f2/f1ratio of 1.2. Responses were recorded at 4, 8, 16, 24 and 32 kHz.

Acoustic Trauma

Animals were exposed to 120 dB octave band noise (8-16 kHz) for 2 hoursin groups of 4 rats, placed in individual compartments of a custom builtcircular cage placed on a 30-cm diameter platform rotating at 3turns/minute (Aqila Innovation, Valbonne, France). The calibrated octaveband noise generated by the RZ6 SigGen software was further amplified bya Crown D-75 amplifier in bridge mode (Crown Audio, Elkhart, Ind., USA)and delivered by four Beyma CP16 compression tweeters (Acustica BeymaS.L., Moncada, Valencia, Spain) positioned 39 cm above the rotatingplatform, each 10 cm from the platform center.

Results Comparative Effect Ofintraperitoneal Administration OfracemicAzasetron, (+)-Azasetron and (−)-Azasetron on Ear Disorder

The ability of the racemic and two enantiomers of azasetronhydrochloride to reduce SSNHL after acoustic trauma was evaluated in arandomized, vehicle-controlled study with 7 weeks old male Wistar rats.Rats were randomized to receive 4.22 mg/kg of the racemic, twoenantiomers of azasetron or vehicle control (physiological saline) byintra-peritoneal injection immediately after acoustic trauma, followedby daily dosing for a total of 14 days. Two days prior to acoustictrauma, baseline audiometric readings using auditory brainstem response(ABR) (8, 16, and 24 kHz) were recorded for each animal. Animals wereexposed to 120-dB octave-band noise (8-16 kHz) for 2 hours to induceacoustic trauma.

Results for auditory threshold shift are provided in FIG. 2 .

Data show that the (+) enantiomer reduced ABR threshold shifts more thanracemic azasetron or (−)-enantiomer across the tested frequencies whencompared to the vehicle control group.

Protective Effect of Oral Administration of (+)-Azasetron Post-Trauma

The ability of the (+)-enantiomer of azasetron hydrochloride to reduceSSNHL after acoustic trauma was evaluated in a randomized,vehicle-controlled study in male Wistar rats, 7 weeks of age. Rats wererandomized to receive 5, 10, or 20 mg/kg of the (+)-enantiomer ofazasetron hydrochloride or vehicle control (physiological saline) byoral gavage immediately after acoustic trauma, followed by daily dosingfor a total of 14 days. Two days prior to acoustic trauma, baselineaudiometric readings using auditory brainstem response (ABR) (8, 16, and24 kHz) and distortion-product otoacoustic emissions (DPOAE) (4, 8, 16,24, and 32 kHz) were recorded for each animal. Post-trauma audiometryrecordings were conducted at 14 days.

Results for auditory threshold shifts are provided in FIG. 3 .

Data show that 10 and 20 mg/kg of the (+) enantiomer significantlyreduced ABR threshold shifts by a mean of 35.1% and 30.5%, respectively,from baseline to day 14, indicating a mean 22.3 and 19.4 dB improvementin hearing, respectively. When compared to the vehicle control group,10- and 20-mg/kg doses of the (+) enantiomer demonstrate a treatmenteffect that reduces the hearing loss (average) from moderately severe(56 to 70 dB) to mild/moderate (26 to 40 dB/41 to 55 dB) based onAmerican Speech Language Hearing Association (ASLHA) criteria (ASLHA,2015).

Otoacoustic emissions were evaluated to further understand the effect ofthe (+) enantiomer on hearing loss. Using animals from the acoustictrauma study, otoacoustic emissions were evaluated followingadministration of 5, 10, or 20 mg/kg of the (+) enantiomer.

The (+) enantiomer (20 mg/kg) significantly reduced DPOAE amplitude lossfrom baseline to day 14 by a mean of 51.2%, suggesting significantlyreduced loss of outer hair cells in the cochlea (FIG. 3 ).

Effect of Oral Administration Ofracemic Azasetron

The ability of the racemic azasetron hydrochloride by oraladministration to reduce SSNHL after acoustic trauma was also evaluated.10 mg/kg of azasetron or vehicle control (physiological saline) wereadministered to rats by oral gavage immediately after acoustic trauma(same protocol as above), followed by daily dosing for a total of 14days.

A small tendency towards a protective effect of the racemic azasetronhydrochloride was observed on ABR thresholds and DPOAE amplitude loss(FIG. 4 ), but unlike treatment with the same dose of (+)-azasetron(FIG. 3 ), it did not reach statistical significance.

Example 3: Activity of (+)-Azasetron Besylate on Cisplatin-InducedHearing Loss and Ototoxicity

The activity of (+)-azasetron besylate to protect against cisplatinototoxicity induced hearing loss was evaluated by assessing effects onABR threshold shift and DPOAE amplitude loss after slow intravenouscisplatin infusion in female Wistar rats (8 mg/kg cisplatin, 30-minuteinfusion). After baseline audiometry, rats were randomly assigned toreceive daily placebo or (+)-azasetron besylate treatment for 14 daysinitiated 15 minutes before cisplatin administration. ABR thresholdshifts and DPOAE amplitude loss were evaluated on day 14 (D14).

In an initial study, animals were divided into 2 groups: Placebo (n=4)and 26.4 mg/kg (+)-azasetron besylate (corresponding to 18.2 mg/kg(+)-azasetron free-base, n=5), both administered by oral gavage for atotal of 14 days. Two days after baseline audiometry (ABR and DPOAEmeasurements), oral treatment was initiated in both groups 15 minutesbefore 8 mg/kg cisplatin infusion during 30 minutes under isofluraneanesthesia. On D14, ABR and DPOAE measurements were performed toevaluate ABR threshold shifts and DPOAE amplitude loss relative tobaseline and the temporal bones of each animal were subsequentlypreserved with PFA fixation for histology (FIG. 5 ).

Daily oral treatment with 26.4 mg/kg significantly reduced both ABRthreshold shifts (˜10-23 dB, up to 79% reduction) (FIG. 5A) and DPOAEamplitude loss (˜3.5-15 dB, up to 78% reduction) (FIG. 5B) at D14 aftercisplatin infusion compared to the placebo treated group.

In a follow-up study with lower doses of (+)-azasetron besylate, animalswere divided into 3 groups: Placebo (n=6), 6.6 mg/kg (+)-azasetronbesylate (corresponding to 5 mg/kg (+)-azasetron free-base, n=7) and13.2 mg/kg (+)-azasetron besylate (corresponding to 10 mg/kg(+)-azasetron free-base, n=5), both administered by oral gavage for atotal of 14 days. Two days after baseline audiometry (ABR and DPOAEmeasurements), oral treatment was initiated in both groups 15 minutesbefore 8 mg/kg cisplatin infusion for 30 minutes under isofluraneanaesthesia. On D14, ABR and DPOAE measurements were performed toevaluate ABR threshold shifts and DPOAE amplitude loss relative tobaseline (FIG. 7 ).

Both 6.6 mg/kg (+)-azasetron besylate (˜23-29 dB, up to 65% reduction)and 13.2 mg/kg (+)-azasetron besylate (22-29 dB, up to 73% reduction)daily oral treatment significantly reduced ABR threshold shifts comparedto placebo (FIG. 7A) and the reduction of DPOAE amplitude loss by 6.6mg/kg (+)-azasetron besylate (1.5-19 dB, up to 78% reduction) alsoreached statistical significance (FIG. 7B).

To evaluate the effect of (+) azasetron on outer hair cells (OHC)survival after cisplatin administration at an ototoxic dose,cochleograms of surviving OHCs were constructed from whole-mount fixedcochleae. In placebo treated animals, 14 days after cisplatinadministration the mean number of surviving OHCs per 200 μm segment haddecreased significantly, starting at 40-60% distance from the apex andfell to −10 cells or less in the basal turn. The number of survivingOHCs was significantly higher in animals having received 14 days of (+)azasetron treatment (p<0.001; FIGS. 6 and 8 ) and while hair cell losswas still observed, it was more gradual throughout the medial and basalturns, with conservation of up to 11-fold more OHCs in the basal turn ofthe cochlea, where the apoptotic effect was most prominent.

These data support the capacity of (+)-azasetron besylate to provide asignificant benefit as a treatment against cisplatin inducedototoxicity, as demonstrated using functional audiometric measures ofhearing (ABR threshold shift reduction, DPOAE amplitude loss reduction)corresponding to recognized clinical outcomes (pure tone audiograms,otoacoustic emissions) supported by histological demonstration ofimproved survival of cochlear outer sensory hair cells in cochleogramsconstructed from fixed tissue samples.

Example 4: Activity of (+)-Azasetron Besylate on Vestibular Dysfunction

The capacity of (+)-azasetron besylate to protect against lesions in theperipheral vestibular system was assessed using the kainate model ofperipheral excitotoxic injury. Following unilateral, transtympanicinjection of the excitotoxic kainic acid leading to the swelling,retraction and uncoupling of synapses between vestibular sensory haircells and primary afferent neurons, baseline recordings of spontaneousnystagmus (pathological eye movements caused by a deficit of thevestibuloculomotor reflex) and postural deviations are performed beforerandomization to treatment groups. Recordings were repeated daily for 3days, then again at D6 and D13 after lesion induction.

Following lesion induction and baseline recordings at t=1 h, femaleLong-Evans were randomized to receive intraperitoneal injections ofplacebo (n=18) or 5.6 mg/kg (+)-azasetron besylate daily for 3 days(t=1/24/25 h). Recording of spontaneous nystagmus and posturaldeviations were repeated at 2/4/6/24/25/48/49 hours and D6/D13 afterinitial lesion induction (FIG. 9 ).

(+)-azasetron besylate treatment significantly improved spontaneousnystagmus after excitotoxic peripheral vestibular lesion (p=0.048, 6-48%improvement), particularly in the early phase before centralcompensation of the vestibulooculomotor reflex deficits, already afterthe first administration. An early as well as lasting significanttreatment effect of (+)-azasetron besylate administration (p<0.001,30-62% improvement) was seen in the reduction of the roll angledeviation (postural head tilt) after excitotoxic peripheral vestibularlesion.

These data demonstrate the treatment benefit of (+)-azasetron besylatefor reducing lesion induced deficits of the peripheral vestibular systemof the inner ear.

1. A method of prevention of a cochlear disease, disorder or conditionin a subject in need thereof, wherein said method comprises theadministration to the subject of a composition comprising (+)-azasetron,of Formula (R)-I:

or a pharmaceutically acceptable salt and/or solvate thereof, and atleast one pharmaceutically acceptable excipient.
 2. The method accordingto claim 1, wherein the composition comprises at least a 90:10 w/wmixture of (+)-azasetron of Formula (R)-I:(−)-azasetron, orpharmaceutically acceptable salts and/or solvates thereof.
 3. The methodaccording to claim 1, wherein the pharmaceutically acceptable salt of(+)-azasetron of Formula (R)-I is selected from the group consisting of:(+)-azasetron besylate, (+)-azasetron malate, and (+)-azasetronhydrochloride.
 4. The method according to claim 1, wherein thecomposition is administered at a dose ranging from about 0.01 mg toabout 100 mg.
 5. The method according to claim 1, wherein thecomposition is administered at a dose ranging from about 20 to about 100mg of free-base equivalent per day.
 6. The method according to claim 1,wherein the composition is administered systemically.
 7. The methodaccording to claim 1, wherein the composition is administered orally. 8.The method according to claim 1, wherein the composition is administeredlocally.
 9. The method according to claim 1, wherein the cochleardisease, disorder or condition is sensorineural hearing loss, cochlearototoxicity, cochlear excitotoxic-inducing occurrence, cochlearhyperacusis, or cochlear tinnitus.
 10. The method according to claim 1,wherein the cochlear disease, disorder or condition is ototoxiccompound-induced hearing loss.
 11. The method according to claim 1,wherein the cochlear disease, disorder or condition is platinum-inducedhearing loss.
 12. The method according to claim 1, wherein the cochleardisease, disorder or condition is cisplatin-induced hearing loss. 13.The method according to claim 1, wherein the subject is diagnosed withcancer.
 14. The method according to claim 1, wherein the subject isdiagnosed with cancer and is awaiting the receipt of, or is receivingplatinum-based chemotherapy.
 15. The method according to claim 1,wherein the subject is diagnosed with cancer and is awaiting the receiptof, or is receiving cisplatin, carboplatin, oxaliplatin or a combinationthereof.
 16. The method according to claim 1, wherein the subject isdiagnosed with cancer and wherein the composition is administeredbefore, during and/or after platinum-based chemotherapy.
 17. The methodaccording to claim 1, wherein the subject is diagnosed with cancer andwherein the composition is administered before, during and/or aftercisplatin-, carboplatin- or oxaliplatin-based chemotherapy or acombination thereof.
 18. The method according to claim 1, wherein thecochlear disease, disorder or condition is sudden sensorineural hearingloss, noise-induced hearing loss, or age-related hearing loss.
 19. Themethod according to claim 1, wherein the composition is an immediaterelease composition or a sustained release composition.
 20. The methodaccording to claim 1, wherein the composition is an orodispersiblecomposition.